The Basidiomycetous Yeasts Cryptococcus diffluens and C. liquefaciens Colonize the Skin of Patients with Atopic Dermatitis

Abstract

Our previous research showed that lipophilic yeasts, Malassezia species, colonize the skin of patients with atopic dermatitis (AD) at a high frequency. In this study, we found that two basidiomycetous yeasts, Cryptococcus diffluens and C. liquefaciens, colonize the skin significantly more frequently in AD patients than in healthy subjects. Transparent dressings were applied to the skin of 36 AD patients and 30 healthy subjects and then transferred onto Sabouraud dextrose agar. Colonies recovered from the medium were identified by DNA sequence analysis of internal transcribed spacer regions and the D1/D2 26S rRNA gene. C. diffluens and C. liquefaciens were isolated from 42% (15/36) and 33% (12/36) of AD patients and from 20% (6/30) and 20% (6/30) of healthy subjects, respectively. In addition, fungal DNA was extracted directly from the dressings and amplified in a specific nested PCR assay. C. diffluens and C. liquefaciens DNA were detected in dressings from 97% (35/36) and 86% (31/36) of the AD patients and 47% (14/30) and 37% (11/30) of the healthy subjects, respectively. These findings show that Malassezia spp. are not the only yeasts that colonize the skin of AD patients; Cryptococcus spp. also are present in a high proportion of patients. The role of these microorganisms in AD is as yet unknown, but the current findings, in combination with previous results, indicate that C. diffluens, C. liquefaciens, M. globosa, and M. restricta together colonize the skin surface of AD patients at a high frequency.

The Role of Sensitization to Malassezia sympodialis in Atopic Eczema

“…Among these the yeasts Cryptococcus diffluens and Cryptococcus liquefaciens colonize the skin of AE patients more frequently than those of healthy subjects [12]; however, a direct involvement of specific IgE against these species has not yet been demonstrated. Coprinus comatus, a basidiomycete involved in the development of respiratory allergies [13] has been proposed as a potential aeroallergen that may provoke AE [14], although the number of patients enrolled in the study was quite low.…”

Occurrence and dominance of yeast species in naturally fermented milk from the Tibetan Plateau of China

“…The primers for the amplification were NL1 (5'-GCATATCAA-TAAGCGGAGGAAAAG-3', forward) and NL4 (5'-GGTCCGTTGTTTCAAGACGG-3', reverse). The nucleotide sequences used are the universal primers for the D1/D2 variable region of the 26S rDNA gene reported by Sugita et al (2003). PCR amplification was performed in a MJ Research RTC-200 thermocycler (Biotech International, Perth, Australia) under the following conditions: initial denaturizing at 94 8C for 5 min, followed by 35 cycles of 94 8C for 60 s, 58 8C for 60 s, 72 8C for 120 s, and a final extension at 72 8C for 10 min.…”

Diversity of lactic acid bacteria and yeasts in Airag and Tarag, traditional fermented milk products of Mongolia

“…The D1/D2 variable region of 26S rRNA gene was amplified by PCR with primers NL-1 (Sugita et al 2003) and NL-4 (Sugita et al 2003) (see Table 2). PCR amplification was performed with a model PTC-200 thermal cycler (MJ Research) with PCR buffer containing 1.5 mM MgCl 2, 200 mM of each dNTP, 1 lM of each primer, 1 U Taq DNA polymerase (Takara), and 10 ng of template DNA in a total volume of 30 ll.…”

“…The amplification program consisted of 1 cycle of 94°C for 2 min; 30 cycles of 94°C for 20 s, 55°C for 20 s, and 72°C for 90 s; and finally 1 cycle of 72°C for 3 min. Kurtzman and Robnett (1997) NL-3A GAGACCGATAGCGAACAAG Yeast 18S-26S rDNA ITS region Kurtzman and Robnett (1997) NL-4 GGTCCGTGTTTCAAGACGG Yeast 18S-26S rDNA ITS region Sugita et al (2003) rRNA gene sequencing and phylogenetic analysis…”