Malassezia-Induced Skin Inflammation and Fungal Burden Analysis in a Murine Model

Published: September 29, 2023

Abstract

Source: Sparber, F. et al., Infecting Mice with Malassezia spp. to Study the Fungus-Host Interaction. J. Vis. Exp. (2019)

This video demonstrates Malassezia-induced skin infection in mice. Malassezia oil suspension is applied over the wounded ear, triggering inflammation and swelling of the ear tissue. Later, the ears are dissected, homogenized, and spread on an agar plate for growth. The number of colonies indicates the fungal burden resulting from skin inflammation.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Cultivation of Malassezia under laboratory conditions NOTE: Store all the reagents and media used for this protocol at room temperature (RT, 20 – 25 °C) unless stated otherwise, as the lower temperature can inhibit fungal growth. Prepare the liqu…

Representative Results

Figure 1: In vitro cultivation of Malassezia.   (A) M. sympodialis strain ATCC 42132 grown for 3 days at 30 °C and 180 rpm in liquid mDixon medium (left) next to a control Erlenmeyer flask containing mDixon medium that was not inoculated (right). (B) Colonies of M. sympodialis strain ATCC 42132 on…

Disclosures

The authors have nothing to disclose.

Materials

Agar Sigma-Aldrich A1296-1KG
Attane Isoflurane Piramal Healthcare
Biosaftey cabinet (BSC) Faster Ultra Safe DASIT GROUP TEC 5594 BSL2 certified
Centrifuge Eppendorf 5415D Compatible with 2ml Eppendorf tubes
Dessicated Ox-bile Sigma-Aldrich 70168-100G
Eppendorf Tubes (2 ml) Eppendorf 0030 120.094
Glucose Sigma-Aldrich 49159-5KG
Gylcerol (99 %) Honeywell 10314830
Heating pad Eickenmeyer 648048
Incubator Hereaus B20 Heraeus 412047753 BSL2 certified
Ketasol (100 mg) Graeub AG 6680416
Magentic heating plate MR Hei-Standard Heidolph Instruments 442-1355
Malassezia spp. ATCC 14522, 14521, 42132
Malt extract Sigma-Aldrich 70167-500G
Multiply Biosphere Tubes (200 µl) Sarstedt AG 7084211 Safelock
Native olive oil Commerc. available
Nonidet P40 Axon Lab A1694,0250
Oditest measurment devise Kroeplin S0247 Range 0-5 mm
Oleic Acid Sigma-Aldrich 75090-5ML
Peptone Oxoid LP0037
Petri dishes Sarstedt AG 82.1473
Phosphat buffered salt solution (PBS, 1x) Amimed/Bioconcept 3-05F39
Rompun (2 %) Bayer KP0BFHR
Shaking incubator Infors Minitron Infors BSL2 certified
Spectrometer Jenway 20308 Optical density measurement at 600nm
Spectrometer Cuvettes Greiner Bio-One 613101
Stainless Steel balls (5mm) ABF KU.5G80 1.3541
Syringes 1 ml Sub-Q BD Bioscience 305501
Tissue Lyzer II Quiagen 85300
Transpore Hypoallergic Tape 3M 1527-1
Tween 40 Sigma-Aldrich P1504-100ML
Vitamin A Retinoli Palmitas Eye Cream BAUSCH & LOMB Commerc. available

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Cite This Article
Malassezia-Induced Skin Inflammation and Fungal Burden Analysis in a Murine Model. J. Vis. Exp. (Pending Publication), e21693, doi: (2023).

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