Carbon and nitrogen limitation increase chitosan antifungal activity in Neurospora crassa and fungal human pathogens.
Chitosan permeabilizes plasma membrane and kills sensitive filamentous fungi and yeast. Membrane fluidity and cell energy determine chitosan sensitivity in fungi. A five-fold reduction of both glucose (main carbon (C) source) and nitrogen (N) increased 2-fold Neurospora crassa sensitivity to chitosan. We linked this increase with production of intracellular reactive oxygen species (ROS) and plasma membrane permeabilization. Releasing N. crassa from nutrient limitation reduced chitosan antifungal activity in spite of high ROS intracellular levels. With lactate instead of glucose, C and N limitation increased N. crassa sensitivity to chitosan further (4-fold) than what glucose did. Nutrient limitation also increased sensitivity of filamentous fungi and yeast human pathogens to chitosan. For Fusarium proliferatum, lowering 100-fold C and N content in the growth medium, increased 16-fold chitosan sensitivity. Similar results were found for Candida spp. (including fluconazole resistant strains) and Cryptococcus spp. Severe C and N limitation increased chitosan antifungal activity for all pathogens tested. Chitosan at 100 μg ml(-1) was lethal for most fungal human pathogens tested but non-toxic to HEK293 and COS7 mammalian cell lines. Besides, chitosan increased 90% survival of Galleria mellonella larvae infected with C. albicans. These results are of paramount for developing chitosan as antifungal.
Laboratory of Plant Pathology, Multidisciplinary Institute for Environmental Studies (MIES) Ramon Margalef, Department of Marine Sciences and Applied Biology, University of Alicante, E-03080 Alicante, Spain. Electronic address: federico.lopez@ua.es.
Federico
F
Lopez-Moya
Laboratory of Medical Mycology, Faculty of Medicine, Miguel Hernández University, Sant Joan d'Alacant, Alicante, E-03550, Spain. Electronic address: colom@umh.es.
Maria F
MF
Colom-Valiente
Department of Biotechnology, Immunology Division, University of Alicante, Alicante, E-03080, Spain. Electronic address: pascual.martinez@ua.es.
Pascual
P
Martinez-Peinado
Institute of Neurosciences, UMH-CSIC, Campus de San Juan, Avd. Ramón y Cajal s/n, Sant Joan d'Alacant, E-03550, Alicante, Spain. Electronic address: jesus.martinezl@umh.es.
Jesus E
JE
Martinez-Lopez
Institute of Neurosciences, UMH-CSIC, Campus de San Juan, Avd. Ramón y Cajal s/n, Sant Joan d'Alacant, E-03550, Alicante, Spain. Electronic address: epuelles@umh.es.
Eduardo
E
Puelles
Department of Biotechnology, Immunology Division, University of Alicante, Alicante, E-03080, Spain. Electronic address: josemiguel@ua.es.
Jose M
JM
Sempere-Ortells
Laboratory of Plant Pathology, Multidisciplinary Institute for Environmental Studies (MIES) Ramon Margalef, Department of Marine Sciences and Applied Biology, University of Alicante, E-03080 Alicante, Spain. Electronic address: lv.lopez@ua.es.
Luis V
LV
Lopez-Llorca
D000818:Animals; D000935:Antifungal Agents / Q000494:pharmacology*; D019556:COS Cells; D002175:Candida / Q000187:drug effects / Q000378:metabolism; D002244:Carbon / Q000378:metabolism*; D002462:Cell Membrane / Q000187:drug effects; D002470:Cell Survival; D048271:Chitosan / Q000494:pharmacology*; D002522:Chlorocebus aethiops; D003454:Cryptococcus / Q000187:drug effects / Q000378:metabolism; D003470:Culture Media / Q000737:chemistry; D004847:Epithelial Cells / Q000187:drug effects / Q000502:physiology; D005670:Fusarium / Q000187:drug effects / Q000378:metabolism; D005947:Glucose / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D007773:Lactates / Q000378:metabolism; D050296:Microbial Viability / Q000187:drug effects; D009492:Neurospora crassa / Q000187:drug effects* / Q000378:metabolism*; D009584:Nitrogen / Q000378:metabolism*; D010539:Permeability / Q000187:drug effects; D017382:Reactive Oxygen Species / Q000378:metabolism / Q000633:toxicity
D000935:Antifungal Agents; D003470:Culture Media; D007773:Lactates; D017382:Reactive Oxygen Species; D002244:Carbon; D048271:Chitosan; D005947:Glucose; D009584:Nitrogen
Candida spp.; Fusarium proliferatum; Mammalian cell lines; Membrane permeabilization; Nutrient limitation; ROS