Rim Cultures Research Articles

1258. The Eyes Have It: Investigating a Cluster of Non-lactose Fermenting Gram-Negative Bacilli From Donor Corneal Rim Tissue

BackgroundFollowing corneal transplant, donor corneal rim tissue are sometimes cultured to help predict the risk of post-keratoplasty endophthalmitis. In July 2016, the Infection Control (IC) team was notified by the microbiology laboratory of three donor corneal rim cultures growing non-lactose fermenting (NLF) Gram-negative bacilli, which was unusual for this type of specimen. The IC team initiated an epidemiological outbreak investigation to determine the source of the NLF Gram-negative bacilli.MethodsA 12-month retrospective review of donor corneal rim cultures was performed from July 2015 to July 2016, with continual prospective monitoring of donor corneal rim cultures. The protocols used to prepare corneal donor tissues were reviewed. The standard protocol included flooding the tissue with povidone iodine followed by rinsing with a sterile saline solution and then placement in a sterile container with Opitsol GS (a preservative solution with gentamycin and streptomycin). The sterile saline rinse that was normally used for processing had been on back order and had been replaced with an alternative brand from March 2016 to July 2016. Unopened bottles of the alternative brand of sterile saline fluid and Optisol GS were sent to an outside laboratory for bacterial culture and remaining product was temporarily quarantined.ResultsMicrobiology review revealed seven donor corneal rim cultures positive for NLF Gram-negative bacilli from May to July 2016. Organisms isolated from the donor corneal rim tissue included Achromobacter xylosoxidans (4), Burkholderia cepacia (3), Stenotrophomonas maltophilia (2), and Elizabethkingia meningoseptica (1). Sterility cultures of Opitsol GS demonstrated no growth. Sterility cultures of the sterile saline rinse grew Gram-positive and -negative bacteria from all samples. A FDA MedWatch was submitted in July 2016, and on September 6, 2016 an FDA recall notice was published. The quarantined saline was permanently removed. No clinical infections associated with the positive donor corneal rim cultures were identified.ConclusionMicrobiologists are the front line for IC surveillance. Close partnership between the IC team and the microbiology laboratory can help identify potential outbreaks by alerting them of the growth of atypical organisms or clusters.Disclosures All authors: No reported disclosures.

Reappraisal of the suitability of corneas from bacteremic donors for use in corneal transplants

BackgroundThis study examined whether corneas from bacteremic donors could be used for corneal transplant.MethodsCorneas donated to the National Eye Bank of Taiwan between 1 January 2014 and 30 June 2017...

Interface Fungal Keratitis After Descemet Stripping Automated Endothelial Keratoplasty: A Review of the Literature With a Focus on Outcomes.

To examine the management and associated outcomes of reported cases of interface fungal infection after Descemet stripping automated endothelial keratoplasty (EK). A literature review identified reports of interface fungal infection after Descemet stripping automated EK. Data retrieved from the same included patient demographics, surgical procedure undertaken, donor rim culture result, clinical course, and outcome. A review of the relevant literature was also conducted. Twenty-four cases of interface fungal infection after Descemet stripping automated EK were analyzed. Mean age of those affected was 66 years with range 52 to 85 years. Interface fungal infection after Descemet stripping automated EK was noted at a mean of 49.2 days postoperatively. Candida species were the causative infection in 79% cases. Penetrating keratoplasty was eventually performed in 79% of cases. The mean posttreatment best-corrected visual acuity was 0.44 logMAR. Penetrating keratoplasty is frequently required to eliminate interface fungal infection after Descemet stripping automated EK. A satisfactory visual outcome is usually achieved.

Candida albicans endophthalmitis following penetrating keratoplasty: A case report

Endophthalmitis is one of the most serious complications of intraocular surgeries, and Candida albicans constitutes the majority of cases of post penetrating keratoplasty (PK) endophthalmitis. The presenting case is a 21-year-old woman with known case of advanced keratoconus and vernal keratoconjunctivitis who underwent PK, and developed recalcitrant anterior uveitis and subsequent whitish lesion on the superonasal of the lens. She was on topical steroids with possible impression of Uretts Zavalia syndrome versus phacoantigenic uveitis following traumatic cataract up to the 50th day of post operation that manifested symptoms of frank endophthalmitis; and underwent deep vitrectomy, lensectomy, localized iridectomy and irrigation with diluted Imipenem and Amphotericin-B. Microbiologic study of a vitreous sample revealed growth of Candida albicans, however optisol culture was negative and donor rim was not evaluated. One week later, intravitreal Amphotericin-B was administered. The patient remained aphakic with clear graft in the 6th month follow up. Due to great correlation of positive donor rim culture with fungal endophthalmitis and the devastating effect of delayed diagnosis, routine donor rim culture is recommended, to aid us in earlier diagnosis in the presence of nonspecific early signs of fungal endophthalmitis.

Clinical features, antimicrobial susceptibilities, and treatment outcomes of patients with culture positive endophthalmitis after penetrating keratoplasty

PurposeTo report the clinical features organisms and treatment outcomes in patients with endophthalmitis after penetrating keratoplasty (PK) MethodsRetrospective noncomparative case series. ResultsEleven eyes of 11 patients with culture positive endophthalmitis after PK were included. The time to diagnosis of endophthalmitis from last PK was less than 1 week in 3/11 (27%), between 1 and 4 weeks in 3/11 (27%), and greater than one month in 5/11 (46%) (range 2–924 days). The distribution of isolates included gram positive (GP) 9/11 (82%), gram negative (GN) 1/11 (9%), and fungal 1/11 (9%) species, respectively. Of GP bacteria tested, 9/9 (100%) were sensitive to Vancomycin. Of fungal isolates tested, none (0/1) were sensitive to Amphoteracin, Fluconazole, and/or Voriconazole. Among patients with rim culture data available, 1/7 (14%) donor rims were culture positive for Candida glabrata and 6/7 (86%) were culture negative. Patients were treated with primary tap and inject in 10/11 (91%) and primary vitrectomy in 1/11 (9%). VA of ≥5/200 was present in 2/11 (18%) at time of endophthalmitis diagnosis, and was recorded in 6/11 (55%) at last follow-up. Conclusions and ImportancePatients with endophthalmitis after PK presented at variable time points after surgery. Gram positive organisms were the most common isolate. VA outcomes after treatment were generally poor.

Association between positive corneal rim cultures and microbiology screening protocols in Ontario

Objective(i) To assess the rate of positive microbiological cultures of corneas prepared by the Eye Bank of Canada (Ontario Division) between January 1, 2012, and December 31, 2013; (ii) to review the microbiology protocols at the 5 major transplant centres in Ontario; and (iii) to assess the incidence of endophthalmitis during the study period. DesignRetrospective chart review. ParticipantsA total of 4186 consecutive cultured corneal tissues prepared by the Eye Bank from January 1, 2012, to December 31, 2013. MethodsRates of culture-positive cornea rims and incidence of postkeratoplasty endophthalmitis at 5 surgical centres in Ontario were determined, and the protocols used to culture rims at each site were concurrently reviewed. Culture results were analyzed via logistic regression for positive cultures. ResultsThe rate of positive cultures at each sites were as follows: centre A, 3.74%; centre B, 3.26%; centre C, 0.51%; centre D, 0.48%; and centre E, 0.04%. Centres A, B, and D were noted to have significantly higher positive rates than centre E. In comparing microbiology protocols, longer incubation period (11 days) was 12 times more likely to be associated with higher positive culture rates than shorter period (4–5 days). Six-month follow-up of all keratoplasties revealed zero reported cases of endophthalmitis. ConclusionsA literature review regarding the predictive value of routine culturing reveals conflicting data. Our findings suggest that differences in the microbiology protocols directly influence the rates of positive rim cultures. Without a standardized protocol, it is not possible to evaluate the predictive value of routine corneal rim culturing in predicting postkeratoplasty endophthalmitis.

Donor cornea tissue in cases of drowning or water submersion: eye banks practice patterns and tissue outcomes.

Surgical use of donor corneal tissue from victims of water submersion (drowning or submersion secondary to death) remains controversial due to limited evidence about the quality of these tissues. To assess the safety of donor corneal tissue from victims of water submersion, an investigation of eye banks' practice patterns and tissue outcomes was conducted. All 79 Eye Bank Association of America accredited eye banks were contacted for a phone interview of practices regarding tissue from victims of water submersion. A retrospective review of corneal tissues from 2014 to 2016 from a large eye bank network was performed to identify all donors submerged in water. Corneal epithelial integrity, endothelial cell density (ECD), rim cultures, and adverse events were analyzed for associations with water submersion characteristics. 49 eye banks (62% response) participated in the survey. 55% of these eye banks had specific, written protocol for tissue eligibility from donors submerged in water. With or without specific protocol, eye banks reported considering water type (84%) and length of time submerged (92%) to determine eligibility. 22% of eye banks reported medical director involvement when eligibility determination was unclear. 79 tissues from 40 donors who were submerged were identified in 2014-2016 eye bank data. No donor tissues had pre-processing corneal infiltrates, positive rim cultures, or adverse events post-keratoplasty. Corneal epithelial integrity and ECD were not associated with water type or length of time submerged. In conclusion, data from a large eye bank network showed no adverse events or outcomes, indicating these tissues may be safe.

Association Between Fungal Contamination and Eye Bank–Prepared Endothelial Keratoplasty Tissue

Fungal contamination and infection from donor tissues processed for endothelial keratoplasty is a growing concern, prompting analysis of donor tissues after processing. To determine whether eyebank-processed endothelial keratoplasty tissue is at higher risk of contamination than unprocessed tissue and to model eyebank processing with regard to room temperature exposure on Candida growth in optisol-gentamicin and streptomycin (GS) with and without antifungal supplementation. An examination of the 2013 Eversight Eyebank Study follow-up database for risk factors associated with post-keratoplasty infection identified an increased risk of positive fungal rim culture results in tissue processed for endothelial keratoplasty vs unprocessed tissue. Processing steps at room temperature were hypothesized as a potential risk factor for promotion of fungal growth between these 2 processes. Candida albicans, Candida glabrata, and Candida parapsilosis endophthalmitis isolates were each inoculated into optisol-GS and subjected to 2 different room temperature incubation regimens reflective of current corneal tissue handling protocols. Eversight Eyebank Study outcomes and measures were follow-up inquiries from 6592 corneal transplants. Efficacy study outcomes and measures were fungal colony-forming units from inoculated vials of optisol-GS taken at 2 different processing temperatures. Donor rim culture results were 3 times more likely to be positive for fungi in endothelial keratoplasty-processed eyes (1.14%) than for other uses (0.37%) (difference, 0.77%; 95% CI, 0.17-.1.37) (P = .009). In vitro, increased room temperature incubation of optisol-GS increased growth of Candida species over time. The addition of caspofungin and voriconazole decreased growth of Candida in a species-dependent manner. Detectable Candida growth in donor rim cultures, associated with a higher rate of post keratoplasty infection, is seen in endothelial keratoplasty tissue vs other uses at the time of transplantation, likely owing in part to eyebank preparation processes extending the time of tissue warming. Reduced room temperature incubation and the addition of antifungal agents decreased growth of Candida species in optisol-GS and should be further explored to reduce the risk of infection.

The value of corneoscleral rim cultures in keratoplasty: a systematic review and cost-effectiveness analysis.

PurposeThis study evaluated the performance of donor corneoscleral rim cultures for predicting infection after corneal transplantation, and determines if there is a correlation between positive corneoscleral rim cultures and postkeratoplasty infection.Design and data sourcesThis was a systematic review, prognostic accuracy analysis, and cost-effectiveness analysis. Databases searched were: Medline (Ovid), Embase (Ovid), CINAHL, Cochrane Library, Web of Science, and BioSis Previews. Grey literature was also explored.Materials and methodsA systematic review was conducted to locate published and unpublished studies. All studies examining corneal button contamination and its association with endophthalmitis and keratitis posttransplantation were included. Extracted data were used to calculate sensitivity, specificity, positive predictive value, and negative predictive value. Cost data from the London Laboratory Services Group in London, ON were used to calculate the cost-effectiveness of culturing donor rim cultures.ResultsOf 7,870 grafts, 954 had a positive rim culture (12.1%), with 12 patients going on to develop keratitis or endophthalmitis (1.3%). The prevalence of keratitis and endophthalmitis in this study was 0.15%, and the positive predictive value 1.5%. Of the 12 infections, nine were fungal and three bacterial. The estimated cost of a positive and negative test result was CAD$45.99 and $14.15, respectively. The cost to run all 7,870 tests was estimated to be $141,735.86, with an incremental cost-effectiveness ratio of $40,215.70.ConclusionThere was a significant divergence between bacterial and fungal rim-culture results. Bacterial cultures predicted clinical infection poorly, did not change management, and were expensive. Fungal cultures predicted clinical infection in over 10% of patients, had the potential to change management, and were 40% less expensive than full rim culturing (bacterial and fungal tests). Fungal rim cultures may be considered in areas where fungal infection rates are high.

Incidence and Outcomes of Positive Donor Corneoscleral Rim Fungal Cultures after Keratoplasty

To determine the incidence of positive corneoscleral donor rim fungal cultures after keratoplasty and to report clinical outcomes of grafts with culture-positive donor rims. Retrospective cohort study. Consecutive donor corneas and keratoplasty recipients at a single tertiary referral center over 20 years. Patient charts were reviewed to determine the incidence of positive donor rim fungal cultures and clinical outcomes of all grafts using contaminated tissue. The primary outcome measures were positive donor rim fungal culture results and the development of postkeratoplasty fungal infection using corresponding corneal tissue. The secondary outcome measure was the impact of postoperative prophylaxis on donor tissue-associated infections. A total of 3414 keratoplasty cases were included in the statistical analysis. Seventy-one cases (2.1%) were associated with a fungal culture-positive donor rim. Candida species were cultured in 40 cases (56.3%). There was a higher incidence of positive rim cultures over the last 5 years of the analytic period compared with the first 15 years (P= 0.018). Fungal keratitis developed in 4 cases (5.6%), and all patients required further surgical intervention to achieve cure. There were no cases of fungal endophthalmitis. Empiric antimycotic prophylaxis initiated at the time of positive culture result reduced the incidence of keratitis from 15.8% in untreated cases to 1.9% in treated cases (P= 0.056). Positive donor rim fungal cultures are uncommon, but carry an unacceptably high risk of postoperative fungal infection. This risk may be reduced with prophylactic antimycotic therapy when culture-positive donor rims are identified.

Impact of Eye Bank Lamellar Tissue Cutting for Endothelial Keratoplasty on Bacterial and Fungal Corneoscleral Donor Rim Cultures After Corneal Transplantation

To determine if the lamellar cut of donor tissue for endothelial keratoplasty (EK) by an eye bank facility is associated with a change in the prevalence of positive bacterial or fungal donor rim cultures after corneal transplantation. A retrospective review was conducted of bacterial and fungal cultures of donor rims used for corneal transplantation at a tertiary eye care center from January 1, 2003, to December 31, 2008, with tissue provided by a single eye bank. The cases were divided into 2 groups. Group 1 ("no-cut") included keratoplasty procedures in which a lamellar cut was not performed. Group 2 ("precut") included EK procedures in which a 4-hour period of prewarming of tissue followed by a lamellar cut was performed in the eye bank before tissue delivery to the operating surgeon. There were 351 donor rim cultures in group 1 and 278 in group 2. Bacterial cultures were positive in 30 donor rims (8.5%) in group 1 and 13 (4.7%) in group 2 (P = 0.058). Positive bacterial cultures were not associated with any postoperative infections. Fungal cultures were positive in 8 donor rims (2.3%) in group 1 and 7 (2.5%) in group 2 (P = 1.0). Positive fungal cultures were associated with 2 cases (13.3%) of postoperative fungal infections. Corneal donor tissue can be precut for EK by trained eye bank personnel without an increased risk of bacterial or fungal contamination.

Historical Consideration of Hwandonghae (the East-Sea Rim) Culture and Cultural Research Method

Historical Consideration of Hwandonghae (the East-Sea Rim) Culture and Cultural Research Method

Conjunctival swabs and corneoscleral rim cultures from corneal transplantation donors as possible early indicators for posttransplant endopthalmitis

To determine by microbiological examinations the rate of conjunctiva and corneoscleral rim contamination of cadaver donor tissues. A prospective review of culture results for 98 cadaver donor conjunctival swabs [mean age 76.4 ± 12.9 years (±SD)]. Transplant patients were evaluated both before and after disinfection with gentamicin. Individual parameters evaluated included detection rates of bacteria, variety of detected bacteria, time interval from donor death to tissue harvest and age of donors. Before disinfection, 60 out of 98 conjunctival swabs exhibited microbial growth, while only 36 out of 98 exhibited growth after disinfection (p = 0.0006). Longer intervals between death and tissue harvest were associated with higher positive microbial growth rates. Prior to disinfection, culture-positive donors (74.1 ± 13.6 years) were significantly younger than culture-negative donors (79.8 ± 10.8 years) (p = 0.024). Positive donor rim cultures were noted in 2 out of the 22 corneal transplantations. Microorganisms isolated from the corneal grafts matched those found in the conjunctiva. It is recommended that the corneoscleral buttons be removed as soon as possible. Cultures of conjunctival swabs collected from donors after disinfection may be useful in determining treatment for postoperative infections occurring after corneal transplantation.

Late-onset Candida Keratitis after Descemet Stripping Automated Endothelial Keratoplasty: Clinical and Confocal Microscopic Report

To report clinical and confocal microscopy features of late-onset Candida albicans keratitis after Descemet stripping automated keratoplasty (DSAEK). We performed clinical and confocal scan on a patient who underwent DSAEK and phacoemulsification for Fuchs endothelial dystrophy and cataract. A 76-year-old woman who underwent uneventful DSAEK and phacoemulsification presented with white to cream deposits 3 months after DSAEK. Confocal microscopy showed a hyperintense deposit at the lenticule and interface. Confocal scan also disclosed intense haze and inflammation at the interface and clusters of hyperreflective round structures resembling epithelial cells within the interface area. No signs of hyphae-like structures were seen. Late-onset symptoms misled us into a bacterial chronic endophthalmitis diagnosis, and the patient was started on topical and systemic antibiotics. Despite intense antibiotic therapy, the patient developed severe endophthalmitis, so we performed anterior vitrectomy and the donor lenticule was removed. Microbiology results from the removed lenticule showed infection by C albicans. Antifungal therapy with systemic and topical voriconazole controlled the infection. Candida interface keratitis is possible after DSAEK. The posterior location of infected tissue poses diagnostic and therapeutic challenges. In our case, the late onset of the symptoms and not performing corneoscleral rim cultures delayed correct diagnosis. This is the first reported case of post-DSAEK Candida keratitis with confocal microscopy images. A hyperintense granular deposit was seen at the lenticule and interface with confocal microscopy. We also observed intense haze, granular round structures resembling epithelial cells, and hyperreflective needle-shaped material at the interface. No hyphae-like structures were seen with confocal imaging.

Interface Fungal Keratitis After Endothelial Keratoplasty: A Clinicopathological Report

The authors report two cases of interface fungal keratitis following Descemet's stripping automated endothelial keratoplasty (DSAEK). Two patients developed culture-proven interface keratitis with Candida albicans and Candida glabrata following DSAEK. Both patients presented with white interface opacities at approximately 1 month after uncomplicated DSAEK. Adjunct confocal microscopy identified fungal elements prior to surgical therapy. A penetrating keratoplasty was performed in both cases after failed medical therapy with fungal elements confirmed by corneal histopathology. Interface fungal keratitis must be recognized as a potential complication of endothelial keratoplasty. Surgeons should consider corneal donor rim cultures on all endothelial keratopathy cases and confocal microscopy on cases with new interface opacities in the early postoperative period. These measures may lead to early identification and treatment of fungal interface infections.

Candida glabrata endophthalmitis following penetrating keratoplasty in a patient with negative donor rim culture

BackgroundCandida glabrata endophthalmitis following keratoplasty is rare and almost always associated with positive donor rim culture.Case presentationA 63-year-old patient, diagnosed Fuch's endothelial dystrophy in both eyes underwent a penetrating keratoplasty in his right eye. He had multiple underlying medical problems, which included diabetes mellitus, hypertension, hypoadrenalism on oral dexamethasone and fatty liver secondary to hypertrigliseridemia. He developed multiple suture abscesses, corneal haziness, retrocorneal white plaques and a level of hypopyon two weeks after an uneventful penetrating keratoplasty in his right eye. Cultures of the donor button and the transport media culture were negative. Candida glabrata was isolated successfully from the aqueous and vitreous taps. He was treated with a combination of topical, intracameral, intravitreal and intravenous Amphotericin B. His final visual acuity remained poor due to the haziness of the corneal button.ConclusionCandida glabrata endophthalmitis following penetrating keratoplasty can occur in negative donor rim and transport media cultures. The growth of the organism is facilitated by the patient's immunocompromised status. Awareness by the ophthalmologists and appropriate choice of antibiotics are mandatory in this challenging condition.

Donor-Related Candida Keratitis After Descemet Stripping Automated Endothelial Keratoplasty

To report 2 cases of donor-to-host transmission of Candida albicans interface keratitis after Descemet stripping automated endothelial keratoplasty (DSAEK). We performed clinical and microbiologic examinations on 2 patients who underwent DSAEK for pseudophakic corneal edema. Two patients underwent uneventful DSAEK surgery using corneal tissue from the same donor. The donor corneoscleral rims were cultured at the time of surgery and both rims subsequently grew C. albicans and Candida glabrata. Approximately 5 weeks after surgery, infiltrates were noted in the DSAEK interface in both of these patients. Despite treatment with antifungal therapy, the Candida keratitis was unable to be controlled medically and required surgical intervention in each case, after which there was no recurrence of infection. Candida interface keratitis can occur after DSAEK. These 2 cases emphasize the importance of donor corneoscleral rim cultures, especially with the increase in lamellar and endothelial keratoplasty, which can make such interface infections more difficult to diagnose and treat. Patients with fungal positive donor corneal rim cultures should be prophylactically treated with antifungal therapy.

Candida Keratitis After Descemet Stripping and Automated Endothelial Keratoplasty

To report the donor-to-host transmission of Candida albicans after Descemet stripping and automated endothelial keratoplasty (DSAEK). An 80-year-old woman with pseudophakic bullous keratopathy developed an infiltrate in the donor corneal lenticule after DSAEK. Donor corneoscleral rim cultures grew C. albicans. Gram stain of the removed corneal lenticule demonstrated budding yeast and pseudohyphae, and cultures yielded C. albicans. Despite topical and systemic antifungal therapy and therapeutic penetrating keratoplasty, the patient developed a blind painful eye and underwent enucleation. This case report indicates that fungal keratitis may occur from donor-to-host transmission after DSAEK. The location of the infected tissue poses diagnostic and therapeutic challenges for the surgeon.

Donor-to-Host Transmission of Candida glabrata to Both Recipients of Corneal Transplants From the Same Donor

To report 2 cases of exogenous Candida glabrata endophthalmitis after penetrating keratoplasty in recipients of corneas from the same donor transplanted on the same day. Case reports with ophthalmologic, electron microscopic, and microbiological findings including fungal strain analysis. Two patients developed fungal keratitis and endophthalmitis caused by the same C. glabrata strain within 1 day after penetrating keratoplasty of corneas from the same donor on the same day. Donor-to-host transmission was postulated when eye bank sterility checks were repeatedly negative. A short death-to-harvesting time, routine donor rim cultures, and respecting of a time interval before transplantation may provide an additional safety feature in dealing with corneal tissue from high-risk donors.

Whole Globe Enucleation Versus In Situ Excision for Donor Corneal Retrieval-A Prospective Comparative Study

The purpose of this study was to compare the results after changing from conventional whole globe enucleation to in situ excision of donor corneas. Donor corneal tissue retrieved by enucleation (n = 50) and in situ excision (n = 50) was quantitatively evaluated prospectively, and the main parameters evaluated were endothelial cell counts, ultrasonic corneal pachymetry, microbial contamination, graft clarity, and postoperative median visual acuity at the end of 3 months. Mean preoperative donor endothelial cell densities were 2174 +/- 123 and 2132 +/- 149 cells per millimeter square after enucleation and in situ excision, respectively (P = 0.13). The preoperative disease distribution and visual potential of recipients were comparable between the 2 groups. Corneal pachymetry was lower in the whole globe group at the end of 3 months (528 microm, enucleation group; and 539 microm, in situ group; P = 0.01). The mean postoperative endothelial cell counts were comparable in both groups at 3 months (1708 +/- 104.8/mm for whole globe group, measured in 40 eyes, vs 1674 +/- 117.4/mm in in situ group, recorded in 39 eyes; P = 0.18). The number of positive corneoscleral rim cultures postkeratoplasty was statistically comparable in both groups (20% in enucleation group and 24% in in situ group; P = 0.62). At 3 months, 98% and 92% of grafts had a graft clarity of >or=3+ in whole globe and in situ groups, respectively (P = 0.16). Seventy-two percent of patients in whole globe group and 67% in in situ group achieved a best-corrected visual acuity of >or=0.1. Postoperative graft infection developed in 2 cases in the in situ group. Changeover from whole globe enucleation technique to in situ excision technique of harvesting donor corneas demonstrated that in situ excision is a viable alternative.