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…
Divulgazioni
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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Citazione di questo articolo
Malassezia-Induced Skin Inflammation and Fungal Burden Analysis in a Murine Model. J. Vis. Exp. (Pending Publication), e21693, doi: (2023).