Effects of Photodynamic Inactivation on the Growth and Antifungal Susceptibility of Rhizopus oryzae

  • Zhaoyang Liu
  • Jing Tang
  • Yi SunEmail author
  • Lujuan GaoEmail author
Original Paper


Mucormycosis is an aggressive and high-mortality opportunistic fungal infection, especially in immunocompromised patients. Conventional antifungals or surgery showed a limited effect on this disease. The antimicrobial photodynamic therapy (aPDT) has been proven to be a promising therapeutic choice against multiple pathogenic fungi. We evaluated the effect of aPDT by using methylene blue (MB) combined with a light emitting diode (LED) on the viability of Rhizopus oryzae, as well as the antifungal susceptibility after aPDT treatment in vitro. A total of six strains were included in this study; MB (8, 16, and 32 μg/ml) was chosen for the photosensitizer, and a light source of LED (635 ± 10 nm, 12 J/cm2) device was used to active it. aPDT with MB (32 μg/ml) and LED was highly effective in cell growth inhibition and exhibited colony-forming unit reductions of up to 4.3log10. The minimal inhibitory concentration ranges of itraconazole, posaconazole, and amphotericin B decreased from > 32 μg/ml to 4–8 μg/ml, 8–16 μg/ml to 0.5–2 μg/ml, and 2–4 μg/ml to 0.25–0.5 μg/ml, respectively, after pre-treatment with MB (8 μg/ml) and LED. In conclusion, aPDT with MB and LED was a promising therapeutic option against R. oryzae infections alone or combined with antifungal agents. However, further investigation is needed to determine the potential for clinic therapy and to elucidate the underlying mechanism.


Photodynamic inactivation Fungi Rhizopus oryzae Antifungal susceptibility 



This work was supported by National Natural Science Foundation of China (31400131 to Lujuan Gao and 81401677 to Yi Sun), and Hubei Province Health and Family Planning Scientific Research Project (WJ2018H178 to Yi Sun).

Compliance with Ethical Standards

Conflict of interest

The authors report no conflicts of interest. The authors alone are responsible for the content and writing of the paper.


  1. 1.
    Bitar D, Van Cauteren D, Lanternier F, Dannaoui E, Che D, Dromer F, Desenclos JC, Lortholary O. Increasing incidence of zygomycosis (mucormycosis), France, 1997–2006. Emerg Infect Dis. 2009;15(9):1395–401.CrossRefGoogle Scholar
  2. 2.
    Roden MM, Zaoutis TE, Buchanan WL, Knudsen TA, Sarkisova TA, Schaufele RL, Sein M, Sein T, Chiou CC, Chu JH, Kontoyiannis DP, Walsh TJ. Epidemiology and outcome of zygomycosis: a review of 929 reported cases. Clin Infect Dis. 2005;41(5):634–53.CrossRefGoogle Scholar
  3. 3.
    Marty FM, Cosimi LA, Baden LR. Breakthrough zygomycosis after voriconazole treatment in recipients of hematopoietic stem-cell transplants. N Engl J Med. 2004;350(9):950–2.CrossRefGoogle Scholar
  4. 4.
    Lewis RE, Kontoyiannis DP. Epidemiology and treatment of mucormycosis. Future Microbiol. 2013;8(9):1163–75.CrossRefGoogle Scholar
  5. 5.
    Husain S, Alexander BD, Munoz P, Avery RK, Houston S, Pruett T, Jacobs R, Dominguez EA, Tollemar JG, Baumgarten K, Yu CM, Wagener MM, Linden P, Kusne S, Singh N. Opportunistic mycelial fungal infections in organ transplant recipients: emerging importance of non-Aspergillus mycelial fungi. Clin Infect Dis. 2003;37(2):221–9.CrossRefGoogle Scholar
  6. 6.
    Qiao J, Li R, Ding Y, Fang H. Photodynamic therapy in the treatment of superficial mycoses: an evidence-based evaluation. Mycopathologia. 2010;170(5):339–43.CrossRefGoogle Scholar
  7. 7.
    Calzavara-Pinton P, Rossi MT, Sala R, Venturini M. Photodynamic antifungal chemotherapy. Photochem Photobiol. 2012;88(3):512–22.CrossRefGoogle Scholar
  8. 8.
    Lu Q, Sun Y, Tian D, Xiang S, Gao L. Effects of Photodynamic Therapy on the Growth and Antifungal Susceptibility of Scedosporium and Lomentospora spp. Mycopathologia. 2017;182(11–12):1037–43.CrossRefGoogle Scholar
  9. 9.
    Gao L, Jiang S, Sun Y, Deng M, Wu Q, Li M, Zeng T. Evaluation of the Effects of Photodynamic Therapy Alone and Combined with Standard Antifungal Therapy on Planktonic Cells and Biofilms of Fusarium spp. and Exophiala spp. Front Microbiol. 2016; 7: 617.Google Scholar
  10. 10.
    Carmello JC, Alves F, Ribeiro AP, Basso FG, de Souza Costa CA, Tedesco AC, Primo FL, Mima EG, Pavarina AC. In vivo photodynamic inactivation of Candida albicans using chloro-aluminum phthalocyanine. Oral Dis. 2016;22(5):415–22.CrossRefGoogle Scholar
  11. 11.
    Hu Y, Huang X, Lu S, Hamblin MR, Mylonakis E, Zhang J, Xi L. Photodynamic therapy combined with terbinafine against chromoblastomycosis and the effect of PDT on Fonsecaea monophora in vitro. Mycopathologia. 2015;179(1–2):103–9.CrossRefGoogle Scholar
  12. 12.
    Nunes Mario DA, Denardi LB, Brayer Pereira DI, Santurio JM, Alves SH. In vitro photodynamic inactivation of Sporothrix schenckii complex species. Med Mycol. 2014;52(7):770–3.CrossRefGoogle Scholar
  13. 13.
    Morton CO, Chau M, Stack C. In vitro combination therapy using low dose clotrimazole and photodynamic therapy leads to enhanced killing of the dermatophyte Trichophyton rubrum. BMC Microbiol. 2014;14:261.CrossRefGoogle Scholar
  14. 14.
    Gilaberte Y, Aspiroz C, Martes MP, Alcalde V, Espinel-Ingroff A, Rezusta A. Treatment of refractory fingernail onychomycosis caused by nondermatophyte molds with methylaminolevulinate photodynamic therapy. J Am Acad Dermatol. 2011;65(3):669–71.CrossRefGoogle Scholar
  15. 15.
    Lyon JP, Pedroso e Silva Azevedo Cde M, Moreira LM, de Lima CJ, de Resende MA. Photodynamic antifungal therapy against chromoblastomycosis. Mycopathologia. 2011;172(4):293–7.CrossRefGoogle Scholar
  16. 16.
    Souza LW, Souza SV, Botelho AC. Distal and lateral toenail onychomycosis caused by Trichophyton rubrum: treatment with photodynamic therapy based on methylene blue dye. An Bras Dermatol. 2014;89(1):184–6.CrossRefGoogle Scholar
  17. 17.
    Clinical and Laboratory Standards Institute. Reference method for broth dilution antifungal susceptibility testing of filamentous fungi; approved standard-2nd ed. CLSI document M38-A2. CLSI, Wayne, PA. 2008.Google Scholar
  18. 18.
    Ribes JA, Vanover-Sams CL, Baker DJ. Zygomycetes in human disease. Clin Microbiol Rev. 2000;13(2):236–301.CrossRefGoogle Scholar
  19. 19.
    Ribeiro AP, Andrade MC, da Silva Jde F, Jorge JH, Primo FL, Tedesco AC, Pavarina AC. Photodynamic inactivation of planktonic cultures and biofilms of Candida albicans mediated by aluminum–chloride–phthalocyanine entrapped in nanoemulsions. Photochem Photobiol. 2013;89(1):111–9.CrossRefGoogle Scholar
  20. 20.
    Carrera ET, Dias HB, Corbi SCT, Marcantonio RAC, Bernardi ACA, Bagnato VS, Hamblin MR, Rastelli ANS. The application of antimicrobial photodynamic therapy (aPDT) in dentistry: a critical review. Laser Phys. 2016;26(12).Google Scholar

Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.Department of Stomatology, Jingzhou Central Hospital, The Second Clinical Medical CollegeYangtze UniversityJingzhouChina
  2. 2.Department of Dermatology, Jingzhou Central Hospital, The Second Clinical Medical CollegeYangtze UniversityJingzhouChina
  3. 3.Department of DermatologyZhongshan Hospital Fudan UniversityShanghaiChina

Personalised recommendations