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Production and Quantification of Virulence Factors in Malassezia Species

Inès Hadrich
Inès Hadrich
, 
Nahed Khemekhem
Nahed Khemekhem
, 
Sourour Neji
Sourour Neji
, 
Houaida Trablesi
Houaida Trablesi
, 
Amin Ilahi
Amin Ilahi
, 
Hayet Sellami
Hayet Sellami
, 
Fattouma Makni
Fattouma Makni
 e 
Ali Ayadi
Ali Ayadi
   | 06 dic 2022
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Article Category: Original Paper
Pubblicato online: 06 dic 2022
Pagine: 529 - 538
Ricevuto: 12 lug 2022
Accettato: 20 set 2022
DOI: https://doi.org/10.33073/pjm-2022-047
Parole chiave
© 2022 Inès Hadrich et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Fig. 1

Different Malassezia species biofilm quantification after crystal violet staining (Abs CV). Error bars represent a standard deviation.
Different Malassezia species biofilm quantification after crystal violet staining (Abs CV). Error bars represent a standard deviation.

Fig. 2

Malassezia species colonies of A) high lipase activity, B) very high lipase activity, C) high phospholipase activity, D) very high phospholipase activity, E) keratinase activity null, F) very high keratinase activity.
Malassezia species colonies of A) high lipase activity, B) very high lipase activity, C) high phospholipase activity, D) very high phospholipase activity, E) keratinase activity null, F) very high keratinase activity.

The sequences of primers and probes used in RT-qPCR for gene expression studies in 20 Malassezia globosa isolates.

Enzymes Names Sequences
Keratinase KerF 5’-ACGTCATGCTCAGATTGCAG-3’
KerR 5’-GACTTCCGCGAAGAACAAAG-3’
KerP 5’-FAM-CACAATTGCTCCCGATACCT-MGB
Lipase LipF 5’-ACCCAACATTTGCTTCGTTC-3’
LipR 5’-TCAATTATCAATGGTCGCGA-3’
LipP 5’-FAM-CCTTGGTGGTCGTCAAAGTT-MGB
Phospholipase PhosF 5’-AACTGGTGGATTTGCTGACC-3’
PhosR 5’-CTTTACGGGTCCAAGGTGAA-3’
PhosP 5’-FAM-CCGTGTGCCATTCTACATTG-MGB
Actin ActF 5’-CTCTCCTTGTACGCCTCTGG-3’
ActR 5’-TTGACAAGATGCTCCGTCAG-3’
ActP 5’-TET-GTATTGTGCTGGACTCGGGT-MGB

Level of phospholipase, lipase, and keratinase production by Malassezia species.

Phospholipase activity number of isolates (rate of isolates) Lipase activity number of isolates (rate of isolates) Keratinase activity number of isolates (rate of isolates)
Mean (Pz) ±SD very high n (%) high n (%) Nul n (%) Mean (Pz) ± SD very high n (%) high n (%) Mean (Pz) ± SD very high n (%) high n (%) null
Malassezia globosa (N = 20 ) 0.64 ±0.03 9(45) 11 (55) 0(0) 0.55±0.02 19 (95) 1(5) 0.65±0.02 10 (50) 6 (30) 4(20)
Malassezia restricta (N = 10) 0.76 ±0.02 0(0) 7(70) 3 (30) 0.36 ±0.03 10 (100) 0(0) 0.69 ±0.02 0(0) 2 (20) 8 (80)
Malassezia furfur (N = 14) 0.64 ±0.02 2 (14.29) 12 (85.71) 0(0) 0.45 ±0.03 14 (100) 0(0) 0.67±0.02 1(7.15) 6 (42.85) 7(50)
Malassezia sympodialis (N = 7) 0.67 ±0.02 1 (14.29) 6 (85.71) 0(0) 0.54 ±0.02 5 (71.43) 2 (28.57) 0.62 ±0.03 1 (14.3) 3 (42.85) 3 (42.85)
Malassezia slooffiae (N = 5) 0.64 ±0.02 2(40) 3 (60) 0(0) 0.4 ±0.04 5(100) 0(0) 0.67±0.02 2(40) 3 (60) 0(0)
Malassezia pachydermatis (N = 21) 0.64 ±0.03 9 (42.86) 12 (57.14) 0(0) 0.31 ±0.02 21 (100) 0(0) 0.68±0.02 3 (14.29) 8 (38.5) 10 (47.66)
Total 0.66 ±0.02 23 (29.87) 51 (66.23) 3(3.9) 0.48 ±0.02 74 (91.1) 3 (3.9) 0.66 ±0.02 17(22.08) 28 (36.36) 32 (41.56)

Enzymatic activities of Malassezia species isolated from different clinical sites.

Folliculitis (N = 12) Neonates group (N = 20) Pityriasis versicolor (N = 10) Control group (N = 35)
Phospholipase activity Mean (Pz) ± SD Malassezia globosa (N = 20) 0.69 ± 0.01 (5/12) – (0/20) 0.74 ± 0.02 (8/10) 0.73 ± 0.02 (7/35)
Malassezia restrict (N = 10) 0.67 ± 0.01 (1/12) – (0/20) 0.8 ± 0.02 (1/10) 0.83 ± 0.02 (8/35)
Malassezia furfur (N = 14) Malassezia sympodialis (N = 7) 0.74 ± 0.01 (6/12) – (0/12) 0.70 ± 0.02 (1/20) – (0/20) 0.69 ± 0.02 (1/10) – (0/10) 0.71 ± 0.01 (6/35) 0.78 ± 0.02 (7/35)
Malassezia slooffiae (N = 5) 0 (0/12) – (0/20) – (0/10) 0.77 ± 0.02 (5/35)
Malassezia pachydermatis(N = 21) 0 (0/12) 0.66 ± 0.02 (19/20) – (0/10) 0.79 ± 0.01 (2/35)
Total 0.70 ± 0.01 0.68 ± 0.02 0.74 ± 0.02 0.77 ± 0.02
Lipase activity Mean (Pz) ± SD Malassezia globosa (N = 20) 0.36 ± 0.01 (5/12) – (0/20) 0.36 ± 0.02 (8/10) 0.38 ± 0.02 (7/35)
Malassezia restricta (N = 10) 0.37 ± 0.01 (1/12) – (0/20) 0.32 ± 0.02 (1/10) 0.36 ± 0.02 (8/35)
Malassezia furfur (N = 14) Malassezia sympodialis (N = 7) 0.32 ± 0.02 (6/12) – (0/12) 0.39 ± 0.02 (1/20) – (0/20) 0.28 ± 0.01 (1/10) – (0/10) 0.36 ± 0.02 (6/35) 0.48 ± 0.02 (7/35)
Malassezia slooffiae (N = 5) – (0/12) – (0/20) – (0/10) 0.4 ± 0.01 (5/35)
Malassezia pachydermatis (N = 21) – (0/12) 0.31 ± 0.01 (19/20) – (0/10) 0.38 ± 0.02 (2/35)
Total 0.35 ± 0.02 0.35 ± 0.02 0.32 ± 0.02 0.39 ± 0.02
Keratinase activity Mean (Pz) ± SD Malassezia globosa (N = 20) 0.75 ± 0.02 (5/12) – (0/20) 0.62 ± 0.02 (8/10) 0.85 ± 0.02 (7/35)
Malassezia restricta (N = 10) 1 (1/12) – (0/20) 0.65 ± 0.02 (1/10) 1 (8/35)
Malassezia furfur (N = 14) Malassezia sympodialis (N = 7) 0.82 ± 0.02 (6/12) – (0/12) 0.53 ± 0.02 (1/20) – (0/20) 0.65 ± 0.02 (1/10) – (0/10) 0.96 ± 0.02 (6/35) 0.84 ± 0.02 (7/35)
Malassezia slooffiae (N = 5) – (0/12) – (0/20) – (0/10) 0.8 ± 0.02 (5/35)
Malassezia pachydermatis (N = 21) – (0/12) 0.85 ± 0.02 (19/20) – (0/10) 0.79 ± 0.02 (2/35)
Total 0.86 ± 0.02 0.69 ± 0.02 0.64 ± 0.02 0.873 ± 0.02

Relative quantification of gene expression and keratinase, lipase, and phospholipase genes copy number in 20 Malassezia globosa strains.

ID Clinical sites RNA relative quantification DNA relative quantification
Keratinase Lipase Phospholipase Keratinase Lipase Phospholipase
S1 Folliculitis 2.11 1.84 2.59 4.43 7.22 3.06
S2 Folliculitis 2.52 7.51 7.57 1.36 2.04 1.08
S3 Folliculitis 3.52 12.99 12.40 1.50 2.12 3.36
S4 Folliculitis 0.81 3.20 3.91 0.32 12.98 0.69
S5 Folliculitis 3.40 5.55 1.48 6.58 9.57 6.36
S6 Pityriasis versicolor 0.49 0.31 0.78 0.29 0.21 0.02
S7 Pityriasis versicolor 2.65 3.52 3.17 1.35 1.46 1.10
S8 Pityriasis versicolor 2.82 2.35 2.54 2.79 7.43 1.34
S9 Pityriasis versicolor 1.25 3.10 2.53 0.77 1.60 0.24
S10 Pityriasis versicolor 2.57 2.55 2.52 1.96 1.56 3.33
S11 Pityriasis versicolor 2.84 2.75 1.32 0.97 1.74 1.03
S12 Pityriasis versicolor 0.96 2.86 3.37 0.46 0.86 2.56
S13 Pityriasis versicolor 2.55 2.86 3.38 1.50 2.58 1.88
S14 Control group 1.53 0.53 0.56 1.42 0.82 0.75
S15 Control group 1.04 1.19 2.55 0.92 1.05 0.86
S16 Control group 0.67 1.36 0.48 0.50 0.61 0.32
S17 Control group 0.94 2.37 2.43 0.50 1.06 2.33
S18 Control group 2.36 0.07 0.48 1.87 0.26 0.49
S19 Control group 0.24 2.52 2.80 0.56 1.31 1.39
S20 Control group 2.51 3.30 1.50 1.31 1.18 0.27

Biofilm production by Malassezia species isolated from different clinical sites.

Groups Malassezia globosa Malassezia restricta Malassezia furfur Malassezia sympodialis Malassezia slooffiae Malassezia pachydermatis Total
Mean Abs ±SD Range Mean Abs ±SD Range Mean Abs ±SD Range Mean Abs ±SD Range Mean Abs ±SD Range Mean Abs ±SD Range Mean Abs ±SD
Folliculitis (N= 12) 0.321 ±0.01 0.155-0.41 0.366 ±0.01 0.301-0.398 0.339 ±0.01 0.317-0.594 0 0 0 0.342±0.01
Neonates (N = 20) group 0 0 0.433 ±0.02 0 0 0.361 ±0.02 0.30-0.54 0.396 ±0.02
Pityriasis versicolor (N = 10) 0.317±0.01 0.16-0.604 0.306 ±0.02 0.316 ±0.02 0 0 0 0.313 ±0.02
Control (N = 35) group 0.328±0.01 0.18-0.51 0.338±0.02 0.12-0.311 0.255 ±0.02 0.12-0.311 0.235±0.02 0.137-0.361 0.256±0.02 0.167-0.331 0.336 ±0.02 0.22-0.44 0.291 ±0.02
Total 0.322 ±0.01 0.155-0.604 0.336 ±0.02 0.12-0.572 0.336 ±0.02 0.12-0.594 0.235±0.02 0.137-0.362 0.256 ±0.03 0.167-0.332 0.349 ±0.02 0.22-0.54
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