ABSTRACT
ABSTRACT A 49-year-old patient with changes in the nails of the hallux for 10 years was diagnosed with onychomycosis. The identity of the causative agent was confirmed as Cladosporium halotolerans from the Cladosporium sphaerospermum species complex using molecular techniques. MALDI-TOF identified the agent as C. sphaerospermum complex species. Overall, species such as onychomycosis agents should attract special attention to avoid mistakes in the identification process while considering a probable contaminant as responsible for the disease. These species deserve attention since there are rare descriptions of them as causes of onychomycosis. It is important to recognize them as causes of disease and not just as a probable contaminant.
ABSTRACT
This study was conducted at the Agriculture College University of Karbala, Iraq to isolate and morphologically and molecularly diagnose thirteen Cladosporium isolates collected from tomato plant residues present in desert regions of Najaf and Karbala provinces, Iraq. We diagnosed the obtained isolates by PCR amplification using the ITS1 and ITS4 universal primer pair followed by sequencing. PCR amplification and analysis of nucleotide sequences using the BLAST program showed that all isolated fungi belong to Cladosporium sphaerospermum. Analysis of the nucleotide sequences of the identified C. sphaerospermum isolates 2, 6, 9, and 10 showed a genetic similarity reached 99%, 98%, 99%, and 99%, respectively, with those previously registered at the National Center for Biotechnology Information (NCBl). By comparing the nucleotide sequences of the identified C. sphaerospermum isolates with the sequences belong to the same fungi and available at NCBI, it was revealed that the identified C. sphaerospermum isolates 2, 6, 9, and 10 have a genetic variation with those previously recorded at the National Center for Biotechnology Information (NCBl); therefore, the identified sequences of C. sphaerospermum isolates have been registered in GenBank database (NCBI) under the accession numbers MN896004, MN896107, MN896963, and MN896971, respectively.
Este estudo foi conduzido na Agriculture College University of Karbala, Iraque, para isolar e diagnosticar morfológica e molecularmente treze isolados de Cladosporium coletados de resíduos de plantas de tomate presentes nas regiões desérticas das províncias de Najaf e Karbala, no Iraque. Diagnosticamos os isolados obtidos por amplificação por PCR usando o par de primers universais ITS1 e ITS4 seguido de sequenciamento. A amplificação por PCR e a análise de sequências de nucleotídeos usando o programa BLAST mostraram que todos os fungos isolados pertencem a Cladosporium sphaerospermum. A análise das sequências de nucleotídeos dos isolados 2, 6, 9 e 10 de C. sphaerospermum identificados mostrou similaridade genética de 99%, 98%, 99% e 99%, respectivamente, com aqueles previamente registrados no National Center for Biotechnology Informações (NCBl). Ao comparar as sequências de nucleotídeos dos isolados de C. sphaerospermum identificados com as sequências pertencentes aos mesmos fungos e disponíveis no NCBI, foi revelado que os isolados 2, 6, 9 e 10 de C. sphaerospermum identificados têm variação genética com aqueles anteriormente registrados no National Center for Biotechnology Information (NCBl). Portanto, as sequências identificadas de isolados de C. sphaerospermum foram registradas no banco de dados GenBank (NCBI) sob os números de acesso MN896004, MN896107, MN896963 e MN896971, respectivamente.
Subject(s)
Animals , Citrus/parasitology , Cladosporium/genetics , Polymerase Chain ReactionABSTRACT
Abstract This study was conducted at the Agriculture College University of Karbala, Iraq to isolate and morphologically and molecularly diagnose thirteen Cladosporium isolates collected from tomato plant residues present in desert regions of Najaf and Karbala provinces, Iraq. We diagnosed the obtained isolates by PCR amplification using the ITS1 and ITS4 universal primer pair followed by sequencing. PCR amplification and analysis of nucleotide sequences using the BLAST program showed that all isolated fungi belong to Cladosporium sphaerospermum. Analysis of the nucleotide sequences of the identified C. sphaerospermum isolates 2, 6, 9, and 10 showed a genetic similarity reached 99%, 98%, 99%, and 99%, respectively, with those previously registered at the National Center for Biotechnology Information (NCBl). By comparing the nucleotide sequences of the identified C. sphaerospermum isolates with the sequences belong to the same fungi and available at NCBI, it was revealed that the identified C. sphaerospermum isolates 2, 6, 9, and 10 have a genetic variation with those previously recorded at the National Center for Biotechnology Information (NCBl); therefore, the identified sequences of C. sphaerospermum isolates have been registered in GenBank database (NCBI) under the accession numbers MN896004, MN896107, MN896963, and MN896971, respectively.
Resumo Este estudo foi conduzido na Agriculture College University of Karbala, Iraque, para isolar e diagnosticar morfológica e molecularmente treze isolados de Cladosporium coletados de resíduos de plantas de tomate presentes nas regiões desérticas das províncias de Najaf e Karbala, no Iraque. Diagnosticamos os isolados obtidos por amplificação por PCR usando o par de primers universais ITS1 e ITS4 seguido de sequenciamento. A amplificação por PCR e a análise de sequências de nucleotídeos usando o programa BLAST mostraram que todos os fungos isolados pertencem a Cladosporium sphaerospermum. A análise das sequências de nucleotídeos dos isolados 2, 6, 9 e 10 de C. sphaerospermum identificados mostrou similaridade genética de 99%, 98%, 99% e 99%, respectivamente, com aqueles previamente registrados no National Center for Biotechnology Informações (NCBl). Ao comparar as sequências de nucleotídeos dos isolados de C. sphaerospermum identificados com as sequências pertencentes aos mesmos fungos e disponíveis no NCBI, foi revelado que os isolados 2, 6, 9 e 10 de C. sphaerospermum identificados têm variação genética com aqueles anteriormente registrados no National Center for Biotechnology Information (NCBl). Portanto, as sequências identificadas de isolados de C. sphaerospermum foram registradas no banco de dados GenBank (NCBI) sob os números de acesso MN896004, MN896107, MN896963 e MN896971, respectivamente.
ABSTRACT
This study was conducted at the Agriculture College University of Karbala, Iraq to isolate and morphologically and molecularly diagnose thirteen Cladosporium isolates collected from tomato plant residues present in desert regions of Najaf and Karbala provinces, Iraq. We diagnosed the obtained isolates by PCR amplification using the ITS1 and ITS4 universal primer pair followed by sequencing. PCR amplification and analysis of nucleotide sequences using the BLAST program showed that all isolated fungi belong to Cladosporium sphaerospermum. Analysis of the nucleotide sequences of the identified C. sphaerospermum isolates 2, 6, 9, and 10 showed a genetic similarity reached 99%, 98%, 99%, and 99%, respectively, with those previously registered at the National Center for Biotechnology Information (NCBl). By comparing the nucleotide sequences of the identified C. sphaerospermum isolates with the sequences belong to the same fungi and available at NCBI, it was revealed that the identified C. sphaerospermum isolates 2, 6, 9, and 10 have a genetic variation with those previously recorded at the National Center for Biotechnology Information (NCBl); therefore, the identified sequences of C. sphaerospermum isolates have been registered in GenBank database (NCBI) under the accession numbers MN896004, MN896107, MN896963, and MN896971, respectively.
Este estudo foi conduzido na Agriculture College University of Karbala, Iraque, para isolar e diagnosticar morfológica e molecularmente treze isolados de Cladosporium coletados de resíduos de plantas de tomate presentes nas regiões desérticas das províncias de Najaf e Karbala, no Iraque. Diagnosticamos os isolados obtidos por amplificação por PCR usando o par de primers universais ITS1 e ITS4 seguido de sequenciamento. A amplificação por PCR e a análise de sequências de nucleotídeos usando o programa BLAST mostraram que todos os fungos isolados pertencem a Cladosporium sphaerospermum. A análise das sequências de nucleotídeos dos isolados 2, 6, 9 e 10 de C. sphaerospermum identificados mostrou similaridade genética de 99%, 98%, 99% e 99%, respectivamente, com aqueles previamente registrados no National Center for Biotechnology Informações (NCBl). Ao comparar as sequências de nucleotídeos dos isolados de C. sphaerospermum identificados com as sequências pertencentes aos mesmos fungos e disponíveis no NCBI, foi revelado que os isolados 2, 6, 9 e 10 de C. sphaerospermum identificados têm variação genética com aqueles anteriormente registrados no National Center for Biotechnology Information (NCBl). Portanto, as sequências identificadas de isolados de C. sphaerospermum foram registradas no banco de dados GenBank (NCBI) sob os números de acesso MN896004, MN896107, MN896963 e MN896971, respectivamente.
Subject(s)
Humans , Cladosporium/genetics , Solanum lycopersicum/genetics , Polymerase Chain ReactionABSTRACT
A 53-year-old male patient was admitted to the hospital on March 26, 2006. Ten years prior to the presentation, a small furuncle developed on the dorsum of his right hand, and subsided after 2-week treatment with erythromycin ointment; one month later, a broadbean-sized cutaneous ulcer developed on the dorsum of the same hand. After anti-infective treatment, the ulcer healed while the lesional skin thickened, and long-term topical treatment with compound dexamethasone acetate cream showed no obvious effect Dermatological examination revealed an irregular verrucous plaque measuring 2.5 cm × 4 cm with little exudation on the dorsum of the right hand. KOH preparation of the skin lesion revealed brown spores. Sabouraud's dextrose agar culture grew restricted, velvety and dark green colony, and microscopy revealed branched, globular conidiophores generated by cladospores. DNA sequencing showed that the isolate was different from Cladosporium sphaerospermum (AB100654) by 2 bases in the sequence of D1/D2 region of 26S rDNA, from Cladosporium sphaerospermum (AY625063) by 5 bases in the sequence of internal transcribed spacer 1 and 2 (ITS 1 and ITS 2), but fully consistent with Cladosporium sphaerospermum (AM 176719) in the sequence of ITS region. The isolate was identified as Cladosporium sphaerospermum. Hematoxylin-eosin stain of the lesional tissue revealed granulomatous changes,and PAS stain demonstrated brown spores. A diagnosis of phaeohyphomycosis was made. Antifungal susceptibility testing indicated that the isolate was highly sensitive to itraconazole. The lesion obviously subsided after treatment with oral itraconazole 0.2 g once daily for 8 weeks, but the patient was lost to follow up 2 months later.
ABSTRACT
Cladosporium sphaerospermum, isolated from salt pans was halotolerant. When grown in the presence of salt, the activities of invertase, isocitrate lyase, fructose-1,6 diphosphate aldolase and malate dehydrogenase were found to be increased and that of amylase decreased. Both, enzyme activation as well as an increase in de novo synthesis of enzymes were found to be some of the mechanisms of salt mediated changes. This may be one of the adaptive mechanisms, in halotolerant Cladosporium sphaerospermum.