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1.
Microorganisms ; 12(6)2024 Jun 05.
Article in English | MEDLINE | ID: mdl-38930533

ABSTRACT

In the present study, we compared the genetic variability of fragments from the internal transcribed spacer region (ITS) and the small subunit ribosomal DNA (SSUrDNA) as nuclear markers, in contrast with the ribosomal protein large two (rpl2) loci, placed in the mitochondrion-related organelles (MROs) within and among human fecal samples with Blastocystis. Samples were analyzed using polymerase chain reaction (PCR)-sequencing, phylogenies, and genetics of population structure analyses were performed. In total, 96 sequences were analyzed, i.e., 33 of SSUrDNA, 35 of rpl2, and 28 of ITS. Only three subtypes (STs) were identified, i.e., ST1 (11.4%), ST2 (28.6%), and ST3 (60%); in all cases, kappa indexes were 1, meaning a perfect agreement among ST assignations. The topologies of phylogenetic inferences were similar among them, clustering to each ST in its specific cluster; discrepancies between phylogeny and assignment of STs were not observed. The STRUCTURE v2.3.4 software assigned three subpopulations corresponding to the STs 1-3, respectively. The population indices were consistent with those previously reported by other groups. Our results suggest the potential use of the ITS and rpl2 genes as molecular markers for Blastocystis subtyping as an alternative approach for the study of the genetic diversity observed within and between human isolates of this microorganism.

2.
J Mycol Med ; 33(1): 101352, 2023 Mar.
Article in English | MEDLINE | ID: mdl-36459816

ABSTRACT

Dermatophytes are keratinophilic fungi that cause skin infections in both humans and animals. Recently, the incidence rates of fungal infections associated with Trichophyton spp. have been considered endemic in many locations. The aim of this study was to isolate and characterize Trichophyton spp. from canines and felines. In the present study, screened 442 canine (n = 386) and feline (n = 56) samples for dermatophytes. Among all the samples, ten isolates were identified as Trichophyton spp. based on micro-morphological features. For comparative analysis, we included three human strains of Trichophyton mentagrophytes complex. In vitro susceptibility of antifungal drugs indicated the highest sensitivity except for fluconazole. The canine and human strains were genetically characterized by sequencing three genes: the internal transcribed spacer region of rDNA, translation elongation factor 1- gene, and beta-tubulin. Based on sequence homology and phylogenetic analysis, the ten canine strains belonged to four different species/ genotypes such as T. mentagrophytes genotype VIII (T. indotineae) (n = 5), T. interdigitale (n = 2), T. simii (n = 2) and T. quinckeanum (n = 1). The three human strains used for comparative analysis were identified as T. mentagrophytes genotype VIII (n = 2) and T. benhamiae (n = 1). The study hence indicates that the T. mentagrophytes genotype VIII, considered as an endemic and emerging human pathogenic clone in India, is also the prevalent in animals.


Subject(s)
Cat Diseases , Dog Diseases , Animals , Cats , Dogs , Humans , Phylogeny , Molecular Epidemiology , Sequence Analysis, DNA , Dog Diseases/epidemiology , Trichophyton , DNA, Fungal/genetics
3.
PeerJ ; 6: e4499, 2018.
Article in English | MEDLINE | ID: mdl-29576968

ABSTRACT

In pursuit of developing fast and accurate species-level molecular identification methods, we tested six DNA barcodes, namely ITS2, matK, rbcLa, ITS2+matK, ITS2+rbcLa, matK+rbcLa and ITS2+matK+rbcLa, for their capacity to identify frequently consumed but geographically isolated medicinal species of Fabaceae and Poaceae indigenous to the desert of Cholistan. Data were analysed by BLASTn sequence similarity, pairwise sequence divergence in TAXONDNA, and phylogenetic (neighbour-joining and maximum-likelihood trees) methods. Comparison of six barcode regions showed that ITS2 has the highest number of variable sites (209/360) for tested Fabaceae and (106/365) Poaceae species, the highest species-level identification (40%) in BLASTn procedure, distinct DNA barcoding gap, 100% correct species identification in BM and BCM functions of TAXONDNA, and clear cladding pattern with high nodal support in phylogenetic trees in both families. ITS2+matK+rbcLa followed ITS2 in its species-level identification capacity. The study was concluded with advocating the DNA barcoding as an effective tool for species identification and ITS2 as the best barcode region in identifying medicinal species of Fabaceae and Poaceae. Current research has practical implementation potential in the fields of pharmaco-vigilance, trade of medicinal plants and biodiversity conservation.

4.
Persoonia ; 31: 42-62, 2013 Dec.
Article in English | MEDLINE | ID: mdl-24761034

ABSTRACT

Current literature accepts 17 species in Penicillium section Sclerotiora. Several produce colonies in bright yellow to orange colours and have monoverticillate conidiophores, apart from P. herquei, P. malachiteum and P. nodositatum, which are biverticillate. The focus of this paper is to refine the concepts of the species currently accepted in the section and introduce five new species, named after the Dutch Royal family as P. vanoranjei, P. maximae, P. amaliae, P. alexiae and P. arianeae. Penicillium vanoranjei produces orange (Dutch = oranje) colonies in culture, and is named after Willem-Alexander Claus George Ferdinand, 'Zijne Koninklijke Hoogheid de Prins van Oranje' (translated from Dutch as: 'His Royal Highness the Prince of Orange') and his family, to coincide with his coronation. We review the current taxonomic positions of P. lilacinoechinulatum and P. nodositatum, both currently considered to be synonyms of P. bilaiae. Sequence data generated in this study show that both species are phylogenetically distinct. Penicillium lilacinoechinulatum is closely related to P. amaliae sp. nov., whereas P. nodositatum does not belong to Penicillium sensu stricto. All species were compared morphologically and phylogenetically, based on ß-tubulin and calmodulin DNA data. A table summarising the morphological characters of all species is included, together with photomicrographs and recommended DNA markers for identification.

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