Molecular identification and antifungal susceptibility of clinical fungal isolates from onychomycosis (uncommon and emerging species)

Mycoses ◽  
2018 ◽  
Vol 62 (2) ◽  
pp. 128-143 ◽  
Author(s):  
Iman Haghani ◽  
Masoomeh Shams-Ghahfarokhi ◽  
Abdolhossein Dalimi Asl ◽  
Tahereh Shokohi ◽  
Mohammad Taghi Hedayati
Keyword(s):  
Molecular Identification ◽  
Antifungal Susceptibility ◽  
Fungal Isolates

scholarly journals First report of Coniella castaneicola causing leaf blight on blueberry (Vaccinium virgatum) in China

Plant Disease ◽  
2021 ◽  
Author(s):  
Jiahao Lai ◽  
Guihong Xiong ◽  
Bing Liu ◽  
Weigang Kuang ◽  
Shuilin Song
Keyword(s):  
Molecular Identification ◽  
Morphological Characteristics ◽  
Leaf Blight ◽  
Yield Potential ◽  
Effective Control ◽  
Economic Losses ◽  
Distilled Water ◽  
First Report ◽  
Fungal Isolates ◽  
Blight Disease

Blueberry (Vaccinium virgatum), an economically important small fruit crop, is characterized by its highly nutritive compounds and high content and wide diversity of bioactive compounds (Miller et al. 2019). In September 2020, an unknown leaf blight disease was observed on Rabbiteye blueberry at the Agricultural Science and Technology Park of Jiangxi Agricultural University in Nanchang, China (28°45'51"N, 115°50'52"E). Disease surveys were conducted at that time, the results showed that disease incidence was 90% from a sampled population of 100 plants in the field, and this disease had not been found at other cultivation fields in Nanchang. Leaf blight disease on blueberry caused the leaves to shrivel and curl, or even fall off, which hindered floral bud development and subsequent yield potential. Symptoms of the disease initially appeared as irregular brown spots (1 to 7 mm in diameter) on the leaves, subsequently coalescing to form large irregular taupe lesions (4 to 15 mm in diameter) which became curly. As the disease progressed, irregular grey-brown and blighted lesion ran throughout the leaf lamina from leaf tip to entire leaf sheath and finally caused dieback and even shoot blight. To identify the causal agent, 15 small pieces (5 mm2) of symptomatic leaves were excised from the junction of diseased and healthy tissue, surface-sterilized in 75% ethanol solution for 30 sec and 0.1% mercuric chloride solution for 2 min, rinsed three times with sterile distilled water, and then incubated on potato dextrose agar (PDA) at 28°C for 5-7 days in darkness. Five fungal isolates showing similar morphological characteristics were obtained as pure cultures by single-spore isolation. All fungal colonies on PDA were white with sparse creeping hyphae. Pycnidia were spherical, light brown, and produced numerous conidia. Conidia were 10.60 to 20.12 × 1.98 to 3.11 µm (average 15.27 × 2.52 µm, n = 100), fusiform, sickle-shaped, light brown, without septa. Based on morphological characteristics, the fungal isolates were suspected to be Coniella castaneicola (Cui 2015). To further confirm the identity of this putative pathogen, two representative isolates LGZ2 and LGZ3 were selected for molecular identification. The internal transcribed spacer region (ITS) and large subunit (LSU) were amplified and sequenced using primers ITS1/ITS4 (Peever et al. 2004) and LROR/LR7 (Castlebury and Rossman 2002). The sequences of ITS region (GenBank accession nos. MW672530 and MW856809) showed 100% identity with accessions numbers KF564280 (576/576 bp), MW208111 (544/544 bp), MW208112 (544/544 bp) of C. castaneicola. LSU gene sequences (GenBank accession nos. MW856810 to 11) was 99.85% (1324/1326 bp, 1329/1331 bp) identical to the sequences of C. castaneicola (KY473971, KR232683 to 84). Pathogenicity was tested on three blueberry varieties (‘Rabbiteye’, ‘Double Peak’ and ‘Pink Lemonade’), and four healthy young leaves of a potted blueberry of each variety with and without injury were inoculated with 20 μl suspension of prepared spores (106 conidia/mL) derived from 7-day-old cultures of LGZ2, respectively. In addition, four leaves of each variety with and without injury were sprayed with sterile distilled water as a control, respectively. The experiment was repeated three times, and all plants were incubated in a growth chamber (a 12h light and 12h dark period, 25°C, RH greater than 80%). After 4 days, all the inoculated leaves started showing disease symptoms (large irregular grey-brown lesions) as those observed in the field and there was no difference in severity recorded between the blueberry varieties, whereas the control leaves showed no symptoms. The fungus was reisolated from the inoculated leaves and confirmed as C. castaneicola by morphological and molecular identification, fulfilling Koch’s postulates. To our knowledge, this is the first report of C. castaneicola causing leaf blight on blueberries in China. The discovery of this new disease and the identification of the pathogen will provide useful information for developing effective control strategies, reducing economic losses in blueberry production, and promoting the development of the blueberry industry.

scholarly journals First report of Curvularia plantarum causing leaf spot on sweet potato (Ipomoea batatas) in China

Plant Disease ◽  
2021 ◽  
Author(s):  
Jiahao Lai ◽  
Tongke Liu ◽  
Bing Liu ◽  
Weigang Kuang ◽  
Shuilin Song
Keyword(s):  
Sweet Potato ◽  
Molecular Identification ◽  
Leaf Spot ◽  
Ipomoea Batatas ◽  
Morphological Characteristics ◽  
Distilled Water ◽  
Gene Sequences ◽  
First Report ◽  
Potted Plants ◽  
Fungal Isolates

Sweet potato [Ipomoea batatas (L.) Lam], is an extremely versatile vegetable that possesses high nutritional values. It is also a valuable medicinal plant having anti-cancer, antidiabetic, and anti-inflammatory activities. In July 2020, leaf spot was observed on leaves of sweet potato in Nanchang, China (28°45'51"N, 115°50'52"E), which affected the growth and development of the crop and caused tuberous roots yield losses of 25%. The disease incidence (total number of diseased plants / total number of surveyed plants × 100%) was 57% from a sampled population of 100 plants in the field. Symptomatic plants initially exhibited small, light brown, irregular-shaped spots on the leaves, subsequently coalescing to form large irregular brown lesions and some lesions finally fell off. Fifteen small pieces (each 5 mm2) of symptomatic leaves were excised from the junction of diseased and healthy tissue, surface sterilized in 75% ethanol solution for 30 sec and 0.1% mercuric chloride solution for 2 min, rinsed three times with sterile distilled water and incubated on potato dextrose agar (PDA) plates at 28°C in darkness. A total of seven fungal isolates with similar morphological characteristics were obtained as pure cultures by single-spore isolation. After 5 days of cultivation at 28°C, dark brown or blackish green colonies were observed, which developed brown, thick-walled, simple, or branched, and septate conidiophores. Conidia were 18.28 to 24.91 × 7.46 to 11.69 µm (average 21.27 × 9.48 µm, n = 100) in size, straight or slightly curved, middle cell unequally enlarged, brown to dark brown, apical, and basal cells slightly paler than the middle cells, with three septa. Based on morphological characteristics, the fungal isolates were suspected to be Curvularia plantarum (Raza et al. 2019). To further confirm the identification, three isolates (LGZ1, LGZ4 and LGZ5) were selected for molecular identification. The internal transcribed spacer region (ITS), glyceraldehyde-3-phosphate-dehydrogenase (GAPDH), and translation elongation factor 1-alpha (EF1-α) genes were amplified and sequenced using primers ITS1/ITS4 (Peever et al. 2004), gpd1/gpd2 (Berbee et al. 1999), EF-983F/EF-2218R (Rehner and Buckley 2005), respectively. The sequences of ITS region of the three isolates (accession nos. MW581905, MZ209268, and MZ227555) shared 100% identity with those of C. plantarum (accession nos. MT410571-72, MN044754-55). Their GAPDH gene sequences were identical (accession nos. MZ224017-19) and shared 100% identity with C. plantarum (accession nos. MN264120, MT432926, and MN053037-38). Similarly, EF1-α gene sequences were identical (accession nos. MZ224020-22) and had 100% identity with C. plantarum (accession nos. MT628901, MN263982-83). A maximum likelihood phylogenetic tree was built based on concatenated data from the sequences of ITS, GAPDH, and EF-1α by using MEGA 5. The three isolates LGZ1, LGZ4, and LGZ5 clustered with C. plantarum. The fungus was identified as C. plantarum by combining morphological and molecular characteristics. Pathogenicity tests were conducted by inoculating a conidial suspension (106 conidia/ml) on three healthy potted I. batatas plants (five leaves wounded with sterile needle of each potted plant were inoculated). In addition, fifteen wounded leaves of three potted plants were sprayed with sterile distilled water as a control. All plants were maintained in a climate box (12 h light/dark) at 25°C with 80% relative humidity. All the inoculated leaves started showing light brown flecks after 7 days, whereas the control leaves showed no symptoms. The pathogenicity test was conducted three times. The fungus was reisolated from all infected leaves of potted plants and confirmed as C. plantarum by morphological and molecular identification, fulfilling Koch’s postulates. To our knowledge, this is the first report of C. plantarum causing leaf spot on sweet potato in China. The discovery of this new disease and the identification of the pathogen will contribute to the disease management, provide useful information for reducing economic losses caused by C. plantarum, and lay a foundation for the further research of resistance breeding.

Ocular Fungal Isolates and Antifungal Susceptibility in Northern China

2007 ◽  
Vol 143 (1) ◽  
pp. 131-133.e1 ◽  
Author(s):  
Sun Xuguang ◽  
Wang Zhixin ◽  
Wang Zhiqun ◽  
Luo Shiyun ◽  
Li Ran
Keyword(s):  
Antifungal Susceptibility ◽  
Northern China ◽  
Fungal Isolates

Molecular identification, biofilm formation and antifungal susceptibility of Rhodotorula spp.

2020 ◽  
Vol 47 (11) ◽  
pp. 8903-8909
Author(s):  
Maral Gharaghani ◽  
Simin Taghipour ◽  
Ali Zarei Mahmoudabadi
Keyword(s):  
Molecular Identification ◽  
Biofilm Formation ◽  
Antifungal Susceptibility

Molecular identification and antifungal susceptibility among clinical isolates of dermatophytes in Shiraz, Iran (2017‐2019)

Mycoses ◽  
2020 ◽  
Author(s):  
Zahra Zareshahrabadi ◽  
Alireza Totonchi ◽  
Ali Rezaei‐Matehkolaei ◽  
Macit Ilkit ◽  
Mehdi Ghahartars ◽  
...  
Keyword(s):  
Molecular Identification ◽  
Antifungal Susceptibility ◽  
Clinical Isolates

Molecular Identification and In Vitro Antifungal Susceptibility of Aspergillus Isolates Recovered from Otomycosis Patients in Western China

2020 ◽  
Vol 185 (3) ◽  
pp. 527-535
Author(s):  
LiLi Zhang ◽  
Xiaodong Wang ◽  
Jos Houbraken ◽  
Huan Mei ◽  
Wanqing Liao ◽  
...  
Keyword(s):  
Molecular Identification ◽  
Antifungal Susceptibility ◽  
Western China ◽  
In Vitro Antifungal Susceptibility

scholarly journals Epidemiology and Antifungal Susceptibility of Bloodstream Fungal Isolates in Pediatric Patients: a Spanish Multicenter Prospective Survey

2011 ◽  
Vol 49 (12) ◽  
pp. 4158-4163 ◽  
Author(s):  
J. Peman ◽  
E. Canton ◽  
M. J. Linares-Sicilia ◽  
E. M. Rosello ◽  
N. Borrell ◽  
...  
Keyword(s):  
Pediatric Patients ◽  
Antifungal Susceptibility ◽  
Prospective Survey ◽  
Fungal Isolates

scholarly journals Antimicrobial resistance among fungi from patients with urinary tract infections in Ojo, Lagos, Nigeria

2021 ◽  
Vol 14 (03) ◽  
pp. 254-264
Author(s):  
Dauphin Dighitoghi Moro ◽  
Oluwole Moses David
Keyword(s):  
Urinary Tract ◽  
Amphotericin B ◽  
Urinary Tract Infections ◽  
Fungal Pathogens ◽  
Fungal Infections ◽  
Antifungal Susceptibility ◽  
Candida Spp ◽  
Fungal Isolates ◽  
Vaginal Swabs ◽  
Tract Infections

The incidence of fungal urinary tract infections has risen gradually and has thus constituted a public health challenge. The aim of this study was to determine the prevalence of urinary tract infections by fungi in two health centres in Ojo, Lagos. A total of 200 patients attending the health centers constituting 160 males’ urines and 40 females’ vaginal swabs were recruited for this study. Midstream urine samples and vaginal swabs were aseptically collected and processed using standard mycological techniques. Fungal isolates were identified based on cultural characteristics, lactophenol blue stain, chlamydospore formation, colony colour on CHROM agar Candida medium and API yeast identification. Antifungal susceptibility testing of the isolates was performed by using the Broth dilution and Kirby-Bauer disk diffusion methods using two of the most commonly used antifungal agents. A total of 122 fungal isolates, of which 68 (55.7%) were Candida spp. and 54(44.3%) Aspergillus spp. were recovered. The Candida spp. included 64 (52.5%) C. albicans and 4(3.3%) C. glabrata while Aspergillus spp. included A. flavus, 20(16.4%), A. fumigatus, 24 (19.8%) and A niger, 10(8.2%). The most common fungal pathogens in the urinary tracts of the subjects were Candida albicans and Aspergillus fumigatus. Both C. albicans and A. fumigatus were highly susceptible to both fluconazole and amphotericin B in dimethyl sulphoxide and water (90-100%). Similarly, all Aspergillus spp. were susceptible to both antifungals except A. flavus which showed a slight resistance (10-15%), which appears to be emerging. Both fluconazole and amphotericin B still show high chances of therapeutic efficacy against fungal infections of the urinary tracts.

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