Cluster Analysis of Human and Animal Pathogenic Microsporum Species and Their Teleomorphic States, Arthroderma Species, Based on the DNA Sequences of Nuclear Ribosomal Internal Transcribed Spacer 1

Abstract

Abstract:We performed a cluster analysis of human and animal pathogenic Microsporum species and their teleomorphic states, Arthroderma species, including A. otae-related species (M. canis, M. audouinii, M. distortum, M. equinum, M. langeronii, and M. ferrugineum) and M. gypseum complex (A. fulvum, A. gypseum, and A. incurvatum) using DNA sequences of nuclear ribosomal internal transcribed spacer 1 (ITS1). The dendrogram showed the members of A. otae-related species to be monophyletic and to construct an extremely closely related cluster with a long horizontal branch. This ITS1-homologous group of A. otae was organized in 6 unique genotypes, while sequences of the members of the ITS J-homologous group of M. gypseum complex are more diverse. This ITS I-based database of Microsporum species and their teleomorphic states will provide a useful and reliable species identification system: it is time-saving (takes two to three days), accurate and applicable even to strains with atypical morphological features or in a non-culturable state.Key words: Microsporum species,Arthroderma species, Internal transcribed spacer 1, DNA sequences Microsporum species are important members of the dermatophytes (filamentous fungi, classified as Euascomycetes and consisting of the genera Microsporum, Trichophyton and Epidermophyton), which have the capacity to invade keratinized tissues of humans and other animals to produce an infection, dermatophytosis (9,16,29 Specific DNA sequences of internal transcribed spacer 1 (ITS I) of ribosomal DNA in the human and animal pathogenic Microsporum species and their teleomorphic states, Arthroderma species, were therefore determined and a cluster analysis performed. ITS I is located between the 18S and 5.8S rONA. As reported previously, the variable ITS regions have been proven useful in resolving relationships between close taxonomic relatives (3), and in the field of medical mycology several phylogenetic studies using the ITS I region were recently documented (1,20,27,28). The authors have reported that it is feasible to successfully perform species identification and/or strain typing of Trichophyton species (the major dermatophytes; 2, 19, 22), which are hard to identify from their morphological features, by demonstrating their dendrogram using a base pair sequence Abbreviations: AFLP, amplified fragment length polymorphism; EDTA, ethylenediaminetetraacetic acid; ITS, internal transcribed spacer; peR, polymerase chain reaction; RAPD, random amplificationof polymorphic DNA; SDA, Sabourauddextrose agar.

Molecular and Phenotypic Characterization of Nannizzia (Arthrodermataceae)

“…The study revealed that Nannizzia species had distinct restriction patterns and this matched with subsequent ITS [2] and multilocus studies [3]. Later, phylogenetic studies [55,56] all demonstrated highly concordant topologies of trees, being largely insensitive to strain and taxon sampling effects. Nannizzia aenigmaticum was published separately as an etiologic agent of tinea corporis [13].…”

Study of the ITS region in an atypical isolate and comparison with six species of Microsporum

“…In the present study, we only include a single clinical specimen. Microsporum gypseum is a complex comprising several species ( Arthroderma gypseum and A. incurvatum ) 11 and to our knowledge there are few reports studying this complex by molecular analysis of ITS regions 18–20 . Microsporum canis , M. ferrugineum and Microsporum CHUS‐126‐02 could not be differentiated by means of these procedures.…”

“…[21] studied with PCR fingerprinting and amplified fragment length polymorphism (AFLP) different strains of Microsporum species and concluded that M. canis , M. ferrugineum and M. audouinii have ITS sequences very similar to A. otae and the set of these three species is conspecific. Makimura et al [20] also carry out a cluster analysis based on the ITS region and reported several strains of the A. otae complex (including M. canis and M. ferrugineum ) presented an extremely closely related cluster with a very similar ITS sequence. Future sequencing studies of other DNA regions are necessary.…”

Isolation and Phylogenetic Analysis of Two Thermotolerant, Fermentative Yeast Strains from Liquid Tapé Ketan (Indonesian Rice Wine)

“…This strain also formed primers 18SF1 (5'-AGG TTT CCG TAG GTG AAC CT-3') and 58SR1 (5'-TTC GCT GCG TTC TTC ATC GA-3') as described by Makimura et al (2001). The thermal cycling profile was an initial denaturation step at 94℃ for 5 min, followed by 25 cycles of 94℃ for 1 min, 60℃ for 15 s and 72℃ for 15 s, and a single final extension step at 72℃ for 10 min.…”