scholarly journals Identification of Trichophyton mentagrophytes strains isolated from patients with dermatophytosis

2019 ◽  
pp. 01-07
Author(s):  
Imen Dhib ◽  
Yaacoub A ◽  
Ben Said M ◽  
Fathallah A ◽  
Zemni R
Keyword(s):  
Rflp Analysis ◽  
Trichophyton Mentagrophytes ◽  
Molecular Techniques ◽  
The Body ◽  
Its2 Region ◽  
Direct Microscopic Examination ◽  
Pcr Rflp ◽  
Tunisian Patients ◽  
Mycological Examination

According to epidemiological, clinical and mycological criteria, it has long been admitted that the Trichophyton mentagrophytes species includes two varieties: a zoophilic variety (var. mentagrophytes) and an anthropophilic variety (var. interdigital) that involve the upper and the lower part of the body respectively. The further application of molecular techniques to the characterization of dermatophyte strains showed that this classification is unreliable. The aim of our study was to assess the usefulness of PCR-RFLP (restriction fragment length polymorphism) and sequencing in the characterization of T. mentagrophytes strains taken from Tunisian patients. The study was carried out in 2008 in the laboratory of Parasitology-Mycology of Farhat Hatched hospital, Sousse, Tunisia. A total of 133 strains were isolated from 133 patients addressed to the laboratory for dermatological lesions very evocative of dermatomycosis. Eighty strains were isolated from lesions located on the lower part of the body (onychomycosis, tinea pedis) and 53 strains from the upper part of the body (tinea capitis, tinea corporis). All strains were submitted to mycological examination (direct microscopic examination, and culture on Sabered medium) and further investigated by using RFLP analysis of the PCR amplified ITS1-5.8 s and ITS2 region of the ribosomal DNA and the MvaI restriction enzyme. In addition, 20 strains were further submitted to a sequencing of the ITS1-5.8 s and ITS2 region. On the basis of mycological criteria all strains were diagnosed as T. mentagrophytes. All strains produced the same RFLP pattern and were identified as T. mentagrophytes interdigital regardless of the location of lesions. Out of the 20 sequenced strains, five were found anthropophilic and 15 were zoophilic. In conclusion, all strains provisionally diagnosed as T. mentagrophytes on the basis of mycological criteria were shown to belong to T. interdigital by using PCR-RFLP and sequencing irrespective of the site of lesions. The predominance of zoophilic strains needs further investigation. Keywords: Dermatophytes; Trichophyton mentagrophytes; PCR-RFLP; Sequencing; Phenotypic features

scholarly journals The use of PCR-RFLP to genetically distinguish the morphologically close species: Ceriodaphnia dubia Richard, 1894 and Ceriodaphnia silvestrii Daday, 1902 (Crustacea Cladocera)

2010 ◽  
Vol 70 (1) ◽  
pp. 121-124 ◽  
Author(s):  
MJ. Abreu ◽  
MJ. Santos-Wisniewski ◽  
O. Rocha ◽  
TC. Orlando
Keyword(s):  
Exotic Species ◽  
Native Species ◽  
Rflp Analysis ◽  
Ceriodaphnia Dubia ◽  
Molecular Techniques ◽  
Rrna Genes ◽  
Its2 Region ◽  
State Of Water ◽  
Pcr Rflp ◽  
Rna Genes

The cladocerans are important components of planktonic and benthic freshwater and good indicators of the trophic state of water bodies. The morphological taxonomy of many species of Cladocera is considered complex with minor differences separating some species. Nowadays, molecular techniques provide a powerful tool to identify and classify different taxonomical levels, using mainly ribosomal RNA genes (rRNA) as molecular markers. In the present work we performed PCR-RFLP to separate Ceriodaphnia dubia, an exotic species in Brazil and the native species Ceriodaphnia silvestrii, widely distributed in Brazilian freshwater. The RFLP analysis of the ITS1-5.8S-ITS2 region of rRNA genes showed to be different between C. dubia and C. silvestrii when using enzymes EcoRI, ApaI and SalI. Thus, the ITS1-5.8S-ITS2 region proved to be a useful molecular marker to differentiate the studied Ceriodaphnia species, which makes the task easier of telling apart species that are morphologically very similar. Also, this methodology might be interesting in determining the distribution of the exotic species C. dubia in Brazilian freshwaters, particularly in cases when C. dubia occurs in the absence of C. silvestrii, a particularly difficult task for ecologists who are not taxonomy specialists.

scholarly journals Characterization of Leishmania Species Isolated from Cutaneous Human Samples from Central Region of Syria by RFLP Analysis

2013 ◽  
Vol 2013 ◽  
pp. 1-5 ◽  
Author(s):  
Samar Anis Al-Nahhas ◽  
Rania Magdy Kaldas
Keyword(s):  
Central Region ◽  
Public Health Problem ◽  
Rflp Analysis ◽  
Leishmania Species ◽  
Etiologic Agent ◽  
Direct Microscopic Examination ◽  
Pcr Rflp ◽  
Polymerase Chain ◽  
Rflp Assay

Cutaneous leishmaniasis (CL) is an endemic disease and a public health problem in Hama governorate located in the central region of Syria. The aim of this study was to characterize Leishmania species isolated from human skin samples. A polymerase chain reaction, restriction fragment length polymorphism (PCR-RFLP) assay, was performed on skin lesion material samples from 32 patients with confirmed CL by direct microscopic examination in order to prove its usefulness and efficiency for identification of Leishmania species. Leishmania tropica (L. tropica) is confirmed as an etiologic agent of CL in this area.

Characterization of Fusarium spp. isolates by PCR-RFLP analysis of the intergenic spacer region of the rRNA gene (rDNA)

2006 ◽  
Vol 106 (3) ◽  
pp. 297-306 ◽  
Author(s):  
A. Llorens ◽  
M.J. Hinojo ◽  
R. Mateo ◽  
M.T. González-Jaén ◽  
F.M. Valle-Algarra ◽  
...  
Keyword(s):  
Intergenic Spacer ◽  
Rflp Analysis ◽  
Rrna Gene ◽  
Fusarium Spp ◽  
Intergenic Spacer Region ◽  
Pcr Rflp

scholarly journals Molecular Characterization of Pathogenicity Locus (PaLoc) and tcdC Genetic Diversity Among tcdA+B+ Clostridioides Difficile Clinical Isolates in Tehran, Iran

2020 ◽  
Author(s):  
Mansoor Kodori ◽  
Zohreh Ghalavand ◽  
Abbas Yadegar ◽  
Gita Eslami ◽  
Masoumeh Azimirad ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Molecular Characterization ◽  
Disease Severity ◽  
Genetic Variants ◽  
Rflp Analysis ◽  
Clostridioides Difficile ◽  
Pcr Rflp ◽  
The Common ◽  
Healthcare Associated

Abstract Background: Clostridioides difficile is the main cause of healthcare-associated diarrhea worldwide. It is proposed that certain C. difficile toxinotypes with distinct pathogenicity locus (PaLoc) variants are associated with disease severity and outcomes. Additionally, few studies have described the common C. difficile toxinotypes, and also little is known about the tcdC variants in Iranian isolates. We characterized the toxinotypes and the tcdC genotypes from a collection of Iranian clinical C. difficile tcdA+B+ isolates with known ribotypes (RTs).Methods: Fifty C. difficile isolates with known RTs and carrying the tcdA and tcdB toxin genes were analyzed. Toxinotyping was carried out based on a PCR-RFLP analysis of a 19.6 kb region encompassing the PaLoc. Genetic diversity of the tcdC gene was determined by the sequencing of the gene.Results: Of the 50 C. difficile isolates investigated, five distinct toxinotypes were recognized. Toxinotypes 0 (33/50, 66%) and V (11/50, 22%) were the most frequently found. C. difficile isolates of the toxinotype 0 mostly belonged to RT 001 (12/33, 36.4%), whereas toxinotype V consisted of RT 126 (9/11, 81.8%). The tcdC sequencing showed six variants (35/50, 70%); tcdC-sc3 (24%), tcdC-A (22%), tcdC-sc9 (18%), tcdC-B (2%), tcdC-sc14 (2%), and tcdC-sc15 (2%). The remaining isolates were wild-types (15/50, 30%) in the tcdC gene.Conclusions: The present study demonstrates that the majority of clinical tcdA+B+ isolates of C. difficile frequently harbor tcdC genetic variants. We also found that the RT 001/ toxinotype 0 and the RT 126/ toxinotype V are the most common types among Iranian isolates. Further studies are needed to investigate the putative association of various tcdC genotypes with CDI severity and its recurrence.

Steinernema apuliae sp. n. (Rhabditida: Steinernematidae): a new entomopathogenic nematode from southern Italy

Zootaxa ◽  
2004 ◽  
Vol 460 (1) ◽  
pp. 1 ◽  
Author(s):  
ORESTE TRIGGIANI ◽  
ZDENEK MRÁ»EK ◽  
ALEX REID
Keyword(s):  
First Generation ◽  
Southern Italy ◽  
Entomopathogenic Nematode ◽  
Rflp Analysis ◽  
Tail Length ◽  
Its Region ◽  
The Body ◽  
Steinernema Glaseri ◽  
Pcr Rflp ◽  
H 41

Steinernema apuliae sp. n. has been found in soil samples collected along a saltpan border habitat in southern Italy characterized by a salted silt soil. This species belongs to the long-IJ nematode group represented by Steinernema glaseri (Steiner, 1929) and Steinernema arenarium (Artyukhovsky, 1967) among others. However, it differs from these taxa in some morphometric values such as V%, H%. Females possess asymmetrical, oblique slit vulva, slant vagina and small flap in the vulval opening. These characteristics are more distinct in second generation females which is different from most other steinernematids; the vulva position is behind the mid-body about 57% to 61% of the body length. First-generation females have a conical-like tip bearing 2 to 3 papilla-like protuberances. Male mucron is absent in both generations. Lightly brown spicules have bluntly pointed tip and elongated manubrium. Third-stage infective juveniles are on average over 1000 m long; the position of the excretory pore is posterior (D% 66) and the hyaline layer is less than half the tail length (H% 41 42). Lateral fields are formed by 8 equally distributed ridges. S. apuliae differs from S. glaseri and S. arenarium and is separated by PCR-RFLP analysis of the ITS region. There were no positive cross-breedings among these species.

scholarly journals Detection and characterization of Leptospira spp. isolated from aborted bovine clinical samples

2011 ◽  
Vol 81 (1) ◽  
pp. 21-25 ◽  
Author(s):  
Hassan Momtaz ◽  
Saadat Moshkelani
Keyword(s):  
Public Health Problem ◽  
Rflp Analysis ◽  
Restriction Enzymes ◽  
Clinical Samples ◽  
Dairy Herds ◽  
Important Public Health Problem ◽  
Pcr Rflp ◽  
Beef Herds ◽  
Bovine Abortion

Leptospira is recognized as an important public health problem worldwide, especially in tropical countries, and is a common cause of abortion in dairy and beef herds. The aim of the present study was to detect and characterize Leptospira as the causative agent of abortion in cattle using a PCR-RFLP in Chaharmahal va Bakhtiari and Isfahan provinces, Iran. A total of 220 bovine aborted foetuses and 120 vaginal discharges from an aborted calf were collected from 64 commercial dairy herds. After isolation of 60 Leptospira spp. from samples, RFLP analysis was carried out with HindIII and HaeIII restriction enzymes in reference strains and isolated for characterization. In a total of 340 specimens, 46 (20.9%) and 14 (11.66%) were identified positive for Leptospira spp. from aborted bovine foetuses and vaginal discharges, respectively. The present results also suggest that L. interrogans serovar hardjo has the highest prevalence in the region under study and L. hardjo is a major pathogen causing bovine abortion in Chaharmahal va Bakhtiari and Isfahan provinces of Iran.

scholarly journals Simultaneous dermatophytosis and keratomycosis caused by Trichophyton interdigitale infection: a case report and literature review

2019 ◽  
Vol 19 (1) ◽  
Author(s):  
Mingrui Zhang ◽  
Lanxiang Jiang ◽  
Fuqiu Li ◽  
Yangchun Xu ◽  
Sha Lv ◽  
...  
Keyword(s):  
Infectious Disease ◽  
Microscopic Examination ◽  
Histopathological Examination ◽  
Trichophyton Mentagrophytes ◽  
The Body ◽  
Fungal Culture ◽  
Left Hand ◽  
Loss Of Vision ◽  
Trichophyton Interdigitale ◽  
Direct Microscopic Examination

Abstract Background Dermatophytosis is a fungal infectious disease caused by dermatophytes, which produce protease and keratinase to digest keratin, leading to the colonization, invasion, and infection of the stratum corneum of the skin, hair shafts, and nails. Trichophyton interdigitale belongs to Trichophyton mentagrophytes complex, which is the common pathogen causing dermatophytosis. Fungal keratitis, also called keratomycosis, is an infectious disease of cornea. Case presentation Here, we report a case of simultaneous dermatophytosis and keratomycosis caused by Trichophyton interdigitale. A 67-year-old man presented with extensive erythema all over the body since 4 years ago, fungal infection of left eye for 2 years, and loss of vision in the eye. These symptoms had become aggravated in the last month. Dermatological examinations showed extensive erythematous plaques with clear borders and scales, scattered red papules with ulceration, and scabs throughout the body. Onychomycosis was observed on the nails of left hand, conjunctival infection with secretion and loss of vision were noted in left eye. Hyaline septate hyphae were observed under direct microscopic examination, fungal culture and internal transcribed spacer sequencing revealed T. interdigitale. Histopathological examination suggested infectious granuloma. A diagnosis of dermatophytosis and keratomycosis caused by T. interdigitale with loss of vision in left eye was made. The patient was treated with luliconazole cream (two applications per day) and itraconazole (100 mg, BID, PO). Complete clinical remission was achieved after 1 month. Subsequently, the patient underwent left eye enucleation in the ophthalmology department. Conclusions In the present study, we reported a case of simultaneous dermatophytosis and keratomycosis caused by T. interdigitale, and reviewed the literature on corneal infection caused by Trichophyton. A total of 10 articles with 45 patients were published between 1973 and 2018. The pathogen of 27 patient were identified to species level. There were T. schoenleinii (17), T. mentagrophytes (4), T. verrucosum (3), T. rubrum (1), T. erinacei (1), and T. interdigitale (1). Five patients had corneal trauma, one had contact lens use history. Direct microscopic examination, fungal culture, and analysis of physiological characteristics were the main methods of identification. Early diagnosis and prompt treatment may help improve the management and outcomes.

scholarly journals Molecular Identification and VOMs Characterization of Saccharomyces cerevisiae Strains Isolated from Madeira Region Winery Environments

Processes ◽  
2020 ◽  
Vol 8 (9) ◽  
pp. 1058 ◽  
Author(s):  
Mariangie Castillo ◽  
Emanuel da Silva ◽  
José S. Câmara ◽  
Mahnaz Khadem
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Solid Phase ◽  
Rflp Analysis ◽  
Selective Adaptation ◽  
Internal Transcribed Spacers ◽  
Gas Chromatography Mass Spectrometry ◽  
Pcr Rflp ◽  
Volatile Organic ◽  
Major Chemical

The quality and typical characteristic of wines depends, among other factors, on the volatile organic metabolites (VOMs) that are biosynthesized by yeasts, mainly Saccharomyces cerevisiae species. The yeast strain influences the diversity and proportions of the VOMs produced during the fermentation process, as the genetic predisposition of the strains is a by-product of selective adaptation to the ecosystem. The present work reports the characterization of S. cerevisiae strains isolated from grape must, used in the Demarcated Region of Madeira (DRM) for winemaking. Yeast species were identified by amplification and by restriction fragment length polymorphism (RFLP) analysis of the region 5.8S-internal transcribed spacers (PCR-RFLP of 5.8S-ITS) of ribosomal DNA (rDNA). The strains identification was performed by analyzing the RFLP pattern of mitochondrial DNA (RFLP-mtDNA). The representative strains were selected for the characterization of the volatile profile through headspace solid-phase microextraction (HS-SPME) followed by gas chromatography-mass spectrometry (GC-MS) analysis. A total of 77 VOMs were identified. Higher alcohols, esters, and fatty acids were the major chemical families representing 63%, 16%, and 9%, respectively, in strain A and 54%, 23%, and 15% in strain B. The results indicate the influence of the strain metabolism in the production of VOMs, many of which probably participate in the aroma of the corresponding wines.

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