Japanese Journal of Ophthalmology

, Volume 61, Issue 5, pp 369–377 | Cite as

Pre-banking microbial contamination of donor conjunctiva and storage medium for penetrating keratoplasty

  • Takenori Inomata
  • Koichi Ono
  • Tsuyoshi Matsuba
  • Tina Shiang
  • Antonio Di Zazzo
  • Satoru Nakatani
  • Masahiro Yamaguchi
  • Nobuyuki Ebihara
  • Akira Murakami
Clinical Investigation



The aims of this study were to investigate the incidence of positive donor tissue cultures before transfer to preservation medium (Optisol™-GS) for penetrating keratoplasty, to verify the efficacy of antibiotics contained in Optisol™-GS by examining the drug susceptibility and to assess the relationship between the results of our microbial assessments as well as donor factors and the incidence of contamination.


We conducted a retrospective, cross-sectional study using Juntendo Eye Bank records for all corneal transplantations. Two hundred donor conjunctiva harvestings and storage medium (EP-II®) cultures were performed between July 2008 and June 2011. We analyzed the associations between donor factors (age, gender, history of cataract surgery, death-to-preservation interval, cause of death) and contamination rates using multivariate analysis by the generalized estimating equation model.


We obtained positive bacterial cultures from 154 of the 200 eyes (77.0%). The isolated bacteria were indigenous, such as coagulase-negative Staphylococci, Corynebacterium sp., and methicillin-resistant Staphylococcus aureus (MRSA). There was significant resistance to levofloxacin (18 eyes, 9.0%) and gentamicin (12 eyes, 6.0%), and no vancomycin-resistant bacteria were detected. The donor factors did not correlate with the prevalence of bacterial contamination in our criteria.


Pre-banking microbial assessment allows for microbial detection, bacterial susceptibility and resistance testing. This is useful for developing preservation mediums containing effective spectrum antibiotic agents for high quality control of corneal banking.


Eye banking Bacterial contamination Donor conjunctiva Corneoscleral rim Preservation medium 



We thank Juntendo Eye Bank, Japan Organ Transplant Network and Oyama Lions Club for their cornea donations. The authors thank Dr. Alireza Mashaghi, Dr. Jiaxu Hong and Dr. Masahiro Nakamura for critically reading and editing the manuscript, Dr. Ayako Nakamura for her microbiological insight and Peter Mallen for his assistance in creating the elegant figure.

Conflicts of interest

All authors declared that they have no conflict of interest.


  1. 1.
    Aiken-O’Neill P, Mannis MJ. Summary of comeal transplant activity Eye Bank Association of America. Cornea. 2002;21:1–3.CrossRefPubMedGoogle Scholar
  2. 2.
    Akova YA, Onat M, Koc F, Nurozler A, Duman S. Microbial keratitis following penetrating keratoplasty. Ophthalmic Surg Lasers. 1999;30:449–55.PubMedGoogle Scholar
  3. 3.
    Williams KA, Brereton HM, Coster DJ. Prospects for genetic modulation of corneal graft survival. Eye. 2009;23:1904–9.CrossRefPubMedGoogle Scholar
  4. 4.
    Bates AK, Kirkness CM, Ficker LA, Steele AD, Rice NS. Microbial keratitis after penetrating keratoplasty. Eye. 1990;4(Pt 1):74–8.CrossRefPubMedGoogle Scholar
  5. 5.
    Constantinou M, Jhanji V, Vajpayee RB. Clinical and microbiological profile of post-penetrating keratoplasty infectious keratitis in failed and clear grafts. Am J Ophthalmol. 2013;155(233–7):e2.Google Scholar
  6. 6.
    Tavakkoli H, Sugar J. Microbial keratitis following penetrating keratoplasty. Ophthalmic Surg. 1994;25:356–60.PubMedGoogle Scholar
  7. 7.
    Antonios SR, Cameron JA, Badr IA, Habash NR, Cotter JB. Contamination of donor cornea: postpenetrating keratoplasty endophthalmitis. Cornea. 1991;10:217–20.CrossRefPubMedGoogle Scholar
  8. 8.
    Cameron JA, Badr IA, Miguel Risco J, Abboud E, el Gonnah S. Endophthalmitis cluster from contaminated donor corneas following penetrating keratoplasty. Can J Ophthalmol. 1998;33:8–13.PubMedGoogle Scholar
  9. 9.
    Guss RB, Koenig S, De La Pena W, Marx M, Kaufman HE. Endophthalmitis after penetrating keratoplasty. Am J Ophthalmol. 1983;95:651–8.CrossRefPubMedGoogle Scholar
  10. 10.
    Leveille AS, McMullan FD, Cavanagh HD. Endophthalmitis following penetrating keratoplasty. Ophthalmology. 1983;90:38–9.CrossRefPubMedGoogle Scholar
  11. 11.
    Pardos GJ, Gallagher MA. Microbial contamination of donor eyes. A retrospective study. Arch Ophthalmol. 1982;100:1611–3.CrossRefPubMedGoogle Scholar
  12. 12.
    Taban M, Behrens A, Newcomb RL, Nobe MY, McDonnell PJ. Incidence of acute endophthalmitis following penetrating keratoplasty: a systematic review. Arch Ophthalmol. 2005;123:605–9.CrossRefPubMedGoogle Scholar
  13. 13.
    Wilhelmus KR, Hassan SS. The prognostic role of donor corneoscleral rim cultures in corneal transplantation. Ophthalmology. 2007;114:440–5.CrossRefPubMedGoogle Scholar
  14. 14.
    Wiffen SJ, Weston BC, Maguire LJ, Bourne WM. The value of routine donor corneal rim cultures in penetrating keratoplasty. Arch Ophthalmol. 1997;115:719–24.CrossRefPubMedGoogle Scholar
  15. 15.
    Everts RJ, Fowler WC, Chang DH, Reller LB. Corneoscleral rim cultures: lack of utility and implications for clinical decision-making and infection prevention in the care of patients undergoing corneal transplantation. Cornea. 2001;20:586–9.CrossRefPubMedGoogle Scholar
  16. 16.
    Gomes JA, Dana MR, Dua HS, Goren MB, Laibson PR, Cohen EJ. Positive donor rim culture in penetrating keratoplasty. Cornea. 1995;14:457–62.CrossRefPubMedGoogle Scholar
  17. 17.
    Rehany U, Balut G, Lefler E, Rumelt S. The prevalence and risk factors for donor corneal button contamination and its association with ocular infection after transplantation. Cornea. 2004;23:649–54.CrossRefPubMedGoogle Scholar
  18. 18.
    Hassan SS, Wilhelmus KR, Dahl P, Davis GC, Roberts RT, Ross KW, et al. Infectious disease risk factors of corneal graft donors. Arch Ophthalmol. 2008;126:235–9.CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Matsumoto M, Suzuma K, Miyamura N, Imamura N, Kitaoka T. Conjunctival swabs and corneoscleral rim cultures from corneal transplantation donors as possible early indicators for posttransplant endopthalmitis. Jpn J Ophthalmol. 2011;55:321–6.CrossRefPubMedGoogle Scholar
  20. 20.
    Linke SJ, Fricke OH, Eddy MT, Bednarz J, Druchkiv V, Kaulfers PM, et al. Risk factors for donor cornea contamination: retrospective analysis of 4546 procured corneas in a single eye bank. Cornea. 2013;32:141–8.CrossRefPubMedGoogle Scholar
  21. 21.
    Cornea Donor Study Investigator Group, gal RL, Dontchev M, Beck RW, Mannis MJ, Holland EJ, et al. The effect of donor age on corneal transplantation outcome results of the cornea donor study. Ophthalmology. 2008;115(4):620–626.e6.CrossRefPubMedCentralGoogle Scholar
  22. 22.
    Mahmood MA, Wagoner MD. Penetrating keratoplasty in eyes with keratoconus and vernal keratoconjunctivitis. Cornea. 2000;19:468–70.CrossRefPubMedGoogle Scholar
  23. 23.
    Sugar J, Montoya M, Dontchev M, Tanner JP, Beck R, Gal R, et al. Donor risk factors for graft failure in the cornea donor study. Cornea. 2009;28:981–5.CrossRefPubMedPubMedCentralGoogle Scholar
  24. 24.
    Woodmansey EJ. Intestinal bacteria and ageing. J Appl Microbiol. 2007;102:1178–86.CrossRefPubMedGoogle Scholar
  25. 25.
    Eye Bank Association of America. Medical Standards 2016 2014 [Available from:
  26. 26.
    Institute CaLS. Performance standards for antimicrobial susceptibility testing, 26th informational supplement M100-S26 2016 [Available from:
  27. 27.
    Kaufman HE, Beuerman RW, Steinemann TL, Thompson HW, Varnell ED. Optisol corneal storage medium. Arch Ophthalmol. 1991;109:864–8.CrossRefPubMedGoogle Scholar
  28. 28.
    Lindstrom RL, Kaufman HE, Skelnik DL, Laing RA, Lass JH, Musch DC, et al. Optisol corneal storage medium. Am J Ophthalmol. 1992;114:345–56.CrossRefPubMedGoogle Scholar
  29. 29.
    Lindstrom RL. Advances in corneal preservation. Trans Am Ophthalmol Soc. 1990;88:555–648.PubMedPubMedCentralGoogle Scholar
  30. 30.
    Means TL, Geroski DH, Hadley A, Lynn MJ, Edelhauser HF. Viability of human corneal endothelium following Optisol-GS storage. Arch Ophthalmol. 1995;113:805–9.CrossRefPubMedGoogle Scholar
  31. 31.
    Mindrup EA, Dubbel PA, Doughman DJ. Betadine decontamination of donor globes. Cornea. 1993;12:324–9.CrossRefPubMedGoogle Scholar
  32. 32.
    Murali S, Jambulingam M, Tiru V, Kulanthai LT, Rajagopal R, Padmanaban P, et al. A study on isolation rate and prevalence of drug resistance among microorganisms isolated from multiorgan donor and donor corneal rim along with a report on existence of bla NDM-1 among Indian population. Curr Eye Res. 2012;37:195–203.CrossRefPubMedGoogle Scholar
  33. 33.
    Mathers WD, Lemp MA. Corneal rim cultures. Cornea. 1987;6:231–3.CrossRefPubMedGoogle Scholar
  34. 34.
    Farrell PL, Fan JT, Smith RE, Trousdale MD. Donor cornea bacterial contamination. Cornea. 1991;10:381–6.CrossRefPubMedGoogle Scholar
  35. 35.
    Borderie VM, Laroche L. Microbiologic study of organ-cultured donor corneas. Transplantation. 1998;66:120–3.CrossRefPubMedGoogle Scholar
  36. 36.
    Sandboe FD, Medin W, Bjerknes R. Toxicity of vancomycin on corneal endothelium in rabbits. Acta Ophthalmol Scand. 1998;76:675–8.CrossRefPubMedGoogle Scholar
  37. 37.
    Cunningham WJ, Moffatt SL, Brookes NH, Twohill HC, Pendergrast DG, Stewart JM, et al. The New Zealand National Eye Bank study: trends in the acquisition and storage of corneal tissue over the decade 2000 to 2009. Cornea. 2012;31:538–45.CrossRefPubMedGoogle Scholar
  38. 38.
    Sugar A, Gal RL, Beck RW, Ruedy KJ, Blanton CL, Feder RS, et al. Baseline donor characteristics in the Cornea Donor Study. Cornea. 2005;24:389–96.CrossRefPubMedGoogle Scholar
  39. 39.
    Reddy SC, Paul G. Bacterial flora of conjunctiva after death. Int J Ophthalmol. 2013;6:632–6.PubMedPubMedCentralGoogle Scholar
  40. 40.
    Van Meter WS, Katz DG, White H, Gayheart R. Effect of death-to-preservation time on donor corneal epithelium. Trans Am Ophthalmol Soc. 2005;103:209–22 (discussion 22-4).PubMedPubMedCentralGoogle Scholar

Copyright information

© Japanese Ophthalmological Society 2017

Authors and Affiliations

  • Takenori Inomata
    • 1
  • Koichi Ono
    • 2
  • Tsuyoshi Matsuba
    • 3
  • Tina Shiang
    • 4
  • Antonio Di Zazzo
    • 5
  • Satoru Nakatani
    • 1
  • Masahiro Yamaguchi
    • 1
  • Nobuyuki Ebihara
    • 6
  • Akira Murakami
    • 1
  1. 1.Department of OphthalmologyJuntendo University Faculty of MedicineTokyoJapan
  2. 2.Department of OphthalmologyJuntendo Tokyo Koto Geriatric Medical Center, Juntendo University School of MedicineTokyoJapan
  3. 3.Advanced Research Center for Human SciencesWaseda UniversityTokorozawaJapan
  4. 4.Department of Ophthalmology, Schepens Eye Research Institute, Massachusetts Eye and Ear InfirmaryHarvard Medical SchoolBostonUSA
  5. 5.BioMolecular and Cellular Laboratories in OphthalmologyIRCCS, G.B. Bietti FoundationRomeItaly
  6. 6.Department of OphthalmologyJuntendo University Urayasu HospitalTomiokaJapan

Personalised recommendations