A histological procedure to study fungal infection in the wax moth Galleria mellonella

  • F. Perdoni University of Milan, Italy.
  • M. Falleni University of Milan, Italy.
  • D. Tosi University of Milan, Italy.
  • D, Cirasola University of Milan, Italy.
  • S. Romagnoli San Paolo Hospital, Italy.
  • P. Braidotti San Paolo Hospital, Italy.
  • E. Clementi University of Pavia, Italy.
  • G. Bulfamante University of Milan, Italy.
  • E. Borghi | elisa.borghi@unimi.it University of Milan, Italy.

Abstract

The invertebrate model Galleria mellonella is a widely used factitious host to study the microbial pathogenesis in vivo. However, a specific procedure for the recovery and the processing of the infected tissues, important for a better understanding of the host-pathogen interactions, has not been reported to our knowledge. In the present study we describe a new procedure of fixation and processing of larval tissue that allows studying the larval topographic anatomy and assessing the morphological changes due to the fungal infection. Lepidopteran larvae were infected with Candida albicans strains displaying various biofilm-forming abilities. The whole larvae were then examined for tissue changes by histological techniques. We show that comparing cutting planes, serial transversal sections of paraffin-embedded larva result in better accuracy and information recovering. Using this technique, it was possible to preserve the integrity of G. mellonella internal structures allowing the detailed analysis of morphological differences in different experimental groups (i.e., healthy vs infected larvae). We were also able to study strain-related differences in the pathogenesis of C. albicans by observing the immune response elicited and the invasiveness of two isolates within the larval tissues. In general, by processing the whole larva and optimizing routinely histochemical stainings, it is possible to visualize and analyse infected tissues. Various degrees of pathogenicity (strain- or inoculum-related), and the infection time course can be described in details. Moreover, the host immune response events can be followed throughout the infectious process leading to a comprehensive picture of the studied phenomenon.

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Author Biographies

F. Perdoni, University of Milan
Department of Health Sciences
M. Falleni, University of Milan
Department of Health Sciences
D. Tosi, University of Milan
Department of Health Sciences
D, Cirasola, University of Milan
Specialization School in Microbiology and Virology
S. Romagnoli, San Paolo Hospital
Pathology Unit
P. Braidotti, San Paolo Hospital
Pathology Unit
E. Clementi, University of Pavia
Department of Biology and Biotechnology "Lazzaro Spallanzani"
G. Bulfamante, University of Milan
Department of Health Sciences
E. Borghi, University of Milan
Department of Health Sciences
Published
2014-09-23
Section
Original Papers
Supporting Agencies
European Society of Clinical Microbiology and Infectious Disease (ESCMID RESEARCH GRANT 2013).
Keywords:
Galleria mellonella, Candida albicans, host-pathogen interaction.
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How to Cite
Perdoni, F., Falleni, M., Tosi, D., Cirasola, D., Romagnoli, S., Braidotti, P., Clementi, E., Bulfamante, G., & Borghi, E. (2014). A histological procedure to study fungal infection in the wax moth Galleria mellonella. European Journal of Histochemistry, 58(3). https://doi.org/10.4081/ejh.2014.2428