bluuebunny/crossref_metadata_2025_split huggingface dataset:
P014 Prevalence and antifungal susceptibility of Wickerhamomyces anomalus in a tertiary care center
Authors: Pandey Abhishek , Paul Raees , Kaur Harsimran , Ghosh Anup , Chakrabarti Arunaloke , Rudramurthy Shivaprakash 
Published: September 2022

Abstract

Poster session 1, September 21, 2022, 12:30 PM - 1:30 PM

Objectives
The spectrum of yeast species causing fungemia has been expanding with emergence of many unusual pathogenic species. One such species is Wickerhamomyces anomalus which has been recognized as an important cause of nosocomial fungemia in neonates and pediatric patients. We evaluated antifungal susceptibility and the burden of antifungal resistance in W. anomalus.

Methods
Species identification of the isolates was performed using MALDI-TOF MS. Antifungal susceptibility testing was done according to the CLSI broth microdilution method following the M27- A3 protocol. C. parapsilosis ATCC22019 and C. krusei ATCC6258 were used as quality control strains. The lowest azole or echinocandin concentration inhibiting 50% visible growth and for amphotericin B, 100% growth-inhibition was taken as minimum inhibitory concentration (MIC). As species-specific breakpoints for resistance are not available for W. anomalus, the susceptibility data was interpreted using CLSI breakpoints (M60 1st edition) as surrogate markers of resistance.

Results
A total of 633 W. anomalous isolated over a 5-year study period from January 2016, through December 2019 at our center, were evaluated for susceptibility to amphotericin B, fluconazole, voriconazole, caspofungin, anidulafungin, and micafungin. The majority of W. anomalus was isolated from neonates (57.1%) followed by other pediatric patients (40.1%). Antifungal therapy was administered in 21% of the patients with fluconazole as the commonest antifungal agent (67.3%) followed by amphotericin B, 31.2%, and voriconazole (1.4%). For amphotericin B, 99.6% (602/604) of the isolates exhibited MIC ≤ 2 mg/l. For fluconazole, 81% of the isolates were susceptible while 19% of the isolates exhibited reduced susceptibility of which 3% were resistant, and 16% exhibited susceptible dose-dependent phenotype. Of all the fluconazole-resistant isolates, two (0.3%) isolates were cross-resistant to voriconazole. Among echinocandins, the resistant rate of 2.8%, 1.2%, and 1.6% wase noted in caspofungin, anidulafungin, and micafungin, respectively.

Conclusions
Wickerhamomyces anomalus exhibited intrinsic susceptibility to all the antifungal agents evaluated. A substantial resistance to fluconazole was noted in this species while resistance to echinocandins was low.
P030 Highly slow-release antifungal wound dressing for chronic dermatophytosis
Authors: Asghari-paskiabi Farnoush , Bonakdar Shahin , Razzaghi-Abyaneh Mehdi 
Published: September 2022

Abstract

Poster session 1, September 21, 2022, 12:30 PM - 1:30 PM

Objectives
The aim of this research was to make a biocompatible and affordable nanofibrous wound dressing that is able to release terbinafine at the site of chronic superficial fungal infection over time.

Methods
Polymer solution (10%) of poly (caprolactone) (PCL) was prepared in hexafluoroisopropanol (HFIP). Terbinafine hydrochloride (TFH) was added equal to 5% of PCL weight for drug-loaded samples. Electrospinning was performed with a 27-G-needle equipped syringe at a distance of 15 cm which the injection flow rate of the solution was 0.2 ml/h and a 30 kV voltage was applied. The measurements of drug release were performed with HPLC. Antifungal tests were done on three different fungal species and MTT assay was done based on ISO-10993 on 24 h L929 cells. The drug release was monitored for 144 h in a human body simulated system (incubation at 37°C, shaking by 30 rpm, and passing the drug through a wet Whatman filter into PBS).
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Results
The mean diameter of fibers was obtained at 1262 nm for PCL nanofibers without TFH and 249 nm for PCL nanofibers with TFH. The drug loading for PCL was 83.4%. Antifungal activity of PCL fibers was examined against a dermatophyte (Trichophyton mentagrophytes), a saprophyte (Aspergillus fumigatus), and a yeast (Candida albicans). The terbinafine hydrochloride-contained electrospun PCL fibers inhibited the growth of T. Mentagrophytes and A. Fumigatus but did not inhibit C. albicans growth. None of the samples showed cytotoxic effects after 24 h and 2 weeks.

Conclusions
The diameter of PCL nanofibers with TFH apparently decreased by fives times (P &gt;.05). PCL nanofibers successfully inhibited two important fungal species while no toxicity was observed in MTT assay for its extraction of 2 weeks. They were able to release TFH slowly over time which makes them suitable for the treatment of chronic superficial fungal infections.
P033 Genomic epidemiology of the antifungal-resistant dermatophytosis epidemic, India, 2017-2019
Authors: Bagal Ujwal , Gade Lalitha , Le Ngoc , Bentz Meghan , Berkow Elizabeth , Hurst Steven , Lockhart Shawn , Litvintseva Anastasia , Min Brian , Sexton Joseph , Uhrlaß Silke , Jackson Brendan , Chow Nancy , Chiller Tom , Nenoff Pietro , Verma Shyam 
Published: September 2022

Abstract

Poster session 1, September 21, 2022, 12:30 PM - 1:30 PM

Objective
The epidemic of antifungal-resistant dermatophytosis in India has been reported. These infections are associated with severe morbidity, resistance to oral itraconazole and terbinafine, and the widespread misuse of topical steroids. Trichophyton indotineae has emerged as the predominant causative agent. In this study, we investigated 162 dermatophytosis infections from eight Indian states using genomic sequencing. The primary objective was to determine whether a clonal outbreak strain is responsible for the current epidemic.

Methods
A total of 161 T. indotineae and one T. rubrum isolates from skin scrapings collected from India in 2017-2019 and previously reported were sent to the U.S. Centers for Disease Control and Prevention (CDC) for genomic analysis. After species identification, genomic DNA was extracted and sequenced using Illumina NovaSeq. Single-nucleotide (SNP) analysis was performed using the portable workflow MycoSNP (v0.21). Briefly, MycoSNP prepared the reference genome, performed pre-processing, aligned sample reads to the reference using the BWA (v 0.7.17) alignment algorithm, and called variants using GATK (v4.1.4.1). High-quality SNPs were used for constructing phylogenetic trees using neighbor-joining (NJ) and maximum likelihood (ML) methods. Further, to understand if infections are genetically clustered by state or region, multi-dimensional scaling (MDS) was applied using the ML tree in R.

Result
SNP analysis identified 1259 450 variant sites which were used to construct an NJ and ML tree. The tree topology from both NJ and ML methods showed consensus. All 161 T. indotineae isolates from India clustered together forming a large, well-supported clade. SNP differences between the samples varied from 0-160 SNPs. Historical isolates available at CDC were included as controls and clustered over 40 000 SNPs from the clade comprising isolates from India. The MDS plot revealed that isolates did not cluster by state or region.

Conclusion
Antifungal-resistant dermatophytosis is an emerging threat with cases of chronic, recurrent infection reported from several countries including India. Additionally, the rapid spread of infections involves person-to-person spread. Our results suggest that a clonal outbreak of a T. indotineae strain is circulating in multiple states in India. Current plans are to expand the geographic scope of the study by including over 10 countries from Europe, Middle East, and the Americas. This work will allow the public health community to better understand the emergence and transmission of antifungal-resistant dermatophytosis worldwide.
P044 Demonstration of the yeasticidal efficacy of povidone-iodine-based commercial antiseptic solutions against Candida auris
Authors: Chesnay Adélaïde , Bailly Eric , Evplanov Victor , Favalli Filippo , Desoubeaux Guillaume 
Published: September 2022

Abstract

Poster session 1, September 21, 2022, 12:30 PM - 1:30 PM

Objectives
Candida auris is an emerging yeast pathogen with worldwide distribution and a great propensity for nosocomial spread. Recent reports have warned of the significant emergence of C. auris in several healthcare facilities. In order to stop its nosocomial transmission, use of antiseptics constitutes the first-line lever of action in fighting against C. auris skin colonization. However, little is known about the efficacy of these products, and moreover, no antiseptics are currently registered for use against C. auris.

Material and Methods
This study investigated the in vitro yeasticidal activity of povidone-iodine (Betadine®) against C. auris, and compared the findings to C. albicans and C. glabrata.

Results
In all the samples, the fungal load was substantially reduced by ≥4.2 Log10 colony-forming units, according to the EN standard 1275:2005. Moreover, even when largely diluted, povidone-iodine products still allowed a sustainable decrease of the yeast viability below 0.1%.

Conclusion
Overall, these results support the use of such commercial antiseptics in the context of colonization with this yeast.
P048 Antifungal susceptibility and mechanism of azole resistance in Candida albicans clinical isolates from oropharyngeal candidiasis patients in Iran
Authors: Jahanshiri Zahra 
Published: September 2022

Abstract

Poster session 1, September 21, 2022, 12:30 PM - 1:30 PM

Objectives
Oropharyngeal candidiasis (OPC) is the most frequent opportunistic fungal infection in head and neck cancer patients. This study was done to investigate the azole susceptibility of Candida albicans (C. albicans) from oropharyngeal candidiasis (OPC) patients and to determine the relationship between ERG11 gene mutations in these isolates and azole resistance.

Methods
A total of 324 clinical isolates of Candida species were collected. Identification of the oral clinical samples was determined by culturing on CHROMagar, carbohydrate assimilation and ITS sequencing methods. Azole susceptibility was tested in vitro in microdilution studies. The ERG11 genes of 42 isolates of C. albicans were amplified and sequenced.

Results
Of the 324 isolates collected, 44.75% (145 isolates) were C. albicans. ERG11 gene was sequenced in 42 isolates. In total, 14 missense mutations were detected in ERG11 genes from 42 isolates. Among them, A945C and T495A substitutions were most prevalent and were known to cause fluconazole resistance.

Conclusions
A total of 14 mutations in the ERG11 gene were identified in azole-resistant C. albicans isolates, which indicated a possible relation with the increase in resistance to azole drugs and the recurrence of oropharyngeal candidiasis. Finding more mutations and relevance requires studies with a higher number of samples.
P051 A case of recalcitrant sporotrichosis by infection of Sporothrix globosa
Authors: Jeong Eunhye , Yim Jeongeun , Kwon Hyeongmok , Choi Jongsoo , Shin Donghoon , Kim Jayoung 
Published: September 2022

Abstract

Poster session 1, September 21, 2022, 12:30 PM - 1:30 PM

Objectives
Sporotrichosis is the leading subcutaneous mycosis caused by the Sporothrix (S.) schenckii complex. S. globosa is the causative organism of fixed sporotrichosis in Korea. The preferred regimen of cutaneous sporotrichosis is itraconazole for 3-6 months, however, there were few studies for recalcitrant sporotrichosis.

Methods
In 2018, we performed a histological examination of a patient who suffered sporotrichosis for 3 years and cultured part of the specimen. Despite various regimens for years, improvement and exacerbation were repeated, so we took another skin biopsy and cultured it in 2021. Isolates from the 2018 and 2021 lesions were identified as S. globosa by ribosomal DNA ITS sequencing (GenBank accession number: MH499862 and MH499863). The in vitro antifungal sensitivity tests were performed by broth microdilution method according to CLSI M38-A2 guidelines or Sensititre YeastOne® manufacturer's instructions. They were incubated at 30°C in a non-CO2 incubator for 7 days.

Results
In 2018, histologically, we observed chronic inflammatory granuloma comprising lymphocytes, histiocytes, and giant cells, and several spores with periodic acid-Schiff (PAS) staining. Microscopic findings and ITS sequences of rRNA gene were identical with S. globosa. The antifungal susceptibility profile in 2018 revealed sensitive to terbinafine (0.125 μg/ml), and moderate to high MIC values for amphotericin B (2 μg/ml), itraconazole (&gt;16 μg/ml), voriconazole (&gt;16 μg/ml), and echinocandins (&gt;16 μg/ml). Treatment with terbinafine, itraconazole, or amphotericin B, the skin lesions were partially improved, but were not cured. In 2021, we took another skin biopsy and culture specimen. Histopathological and mycological examination results were the same as before. The antifungal susceptibility profile revealed sensitive to itraconazole (0.5/ml), and high MIC for others. Clinically, skin lesions were not improved with the use of itraconazole 200 mg/d. Itraconazole 400 mg/d with local heating induced moderate improvement. There was no evidence of immune deficiency.

Conclusion
We experienced recalcitrant sporotrichosis that did not respond to itraconazole and terbinafine, and the sensitivity of antifungal was changed. In this case, the combination treatment including local heating, saturated KI may be considered, and frequent antifungal susceptibility tests are needed.
P054 Two effects of curcumin to Candida albicans
Authors: Kajiwara Susumu , Lee Yean Sheng , Widiyanto Tria Widiasih , Chen Xinyue , Orihara Kanami , Shibata Hiroyuki 
Published: September 2022

Abstract

Poster session 1, September 21, 2022, 12:30 PM - 1:30 PM

Objectives
Candida albicans is a pathogenic yeast that causes candidiasis in immunocompromised patients. The overuse of antifungal drugs has led to the development of resistance to such drugs by this fungus, which is a major challenge in antifungal chemotherapy. The utilization of natural products is a significant trial for the development of new antifungals. Curcumin, one such natural product, has been widely studied as a drug candidate and is reported to exhibit antifungal activity against C. albicans. Although studies of the mechanism of curcumin against human cancer cells have shown that it inhibits heat shock protein 90 (Hsp90), little is known about its molecular function against C. albicans. In this work, we investigated the relationship between curcumin and Hsp90 of C. albicans.

Methods
For the molecular genetical analyses of C. albicans Hsp90, a doxycycline-mediated HSP90 strain and a HSP90-overexpressing strain of this fungus were constructed. The effect of curcumin on the gene expression of HSF1, AHR1, HOG1, and CDR1 as well as HSP90 was analyzed. Moreover, the stress responses to high temperature and osmotic pressure and the drug efflux of these strains were investigated.

Results
Curcumin reduced the transcription of HSP90 at the post-transcriptional level and it was suggested to lead to the decrease in Hsp90. This phenomenon resulted in the downregulation of HOG1 and CDR1. In addition, we confirmed curcumin also inhibited Cdr1 efflux activity in C. albicans.

Conclusion
Curcumin was suggested to influence not only HSP90 expression but also Cdr1 activity in C. albicans.
P076 The promising antimycotic activities of a novel cyclic antifungal lipopeptide against Human Dermatophyte Isolates
Authors: Ramesh Swetha , Shaw Dipika , Madduri Madhuri , Rudramurthy Shivaprakash M , Roy Utpal 
Published: September 2022

Abstract

Poster session 1, September 21, 2022, 12:30 PM - 1:30 PM

Objectives
To determine the minimum inhibitory concentrations (MICs) of a novel antifungal lipopeptide against clinical isolates of dermatophytes of human origin.

Methods
To perform antifungal susceptibility testing (AFST) by CLSI microbroth dilution method, a reversed-phase high-performance liquid chromatography (RP-HPLC) purified lipopeptide of 1071.4 Da from a wild-type soil isolate Bacillus subtilis was used and compared with the standard allylamine terbinafine MICs. Briefly, 1200 ml of cell-free culture supernatant was extracted using a solvent mixture and silica gel (230-400 mesh size)-based adsorption chromatography. The semi-preparative RP-HPLC system consisted of an Agilent quaternary pump and a variable wavelength detector equipped with a Phenomenex Luna C18 column (10 mm × 250 mm, 5 μm). The solvent system for RP- HPLC was (A) water with 0.1% trifluoroacetic acid (TFA) and (B) acetonitrile containing 0.1% TFA. The gradient of solvent B used for purification was as follows: 0%-54% for 0-20 min at the flow rate of 1 ml/min, 54%-60% from 20-48 min at 0.5 ml/min, 60%-100% from 48-58 min at 0.5 ml/min, 100%-0% from 58-65 min at 1 ml/min and monitored at 210 nm.

Results
Superficial skin infections are caused by dermatophytes including Trichophyton spp. Nowadays, resistance to terbinafine in Trichophyton spp. isolates with higher MICs have been documented in India. We report here the antifungal prowess of a novel small antifungal lipopeptide against 20 clinical isolates of Trichophyton spp. of human origins (from human skin scrapings and nails) with the clinical diagnosis of tinea corporis/cruris and tinea unguium. The representative photographs of T. tonsurans, T. rubrum, and T. mentagrophytes complex are provided below. A total of 6 isolates of T. mentagrophytes and T. rubrum carry point mutations at F397L of squalene epoxidase (SQLE) protein. The in vitro antifungal efficacies determined by AFST revealed that the lipopeptide showed less or equivalent MICs (100% inhibition) in the case of five dermatophytes. The lipopeptide drug has exhibited improved MICs against two T. mentagrophytes complex and two T. rubrum isolates with amino acid substitution F397L in SQLE protein. Trichophyton mentagrophytes complex and T. rubrum with F397L mutations were inhibited at MICs 4-32 μg/mL of terbinafine. In comparison, the lipopeptide showed 4-16 μg/ml (100% inhibition). In the case of all four Trichophyton tonsurans, the MICs ranged between 2-4 μg/ml for the lipopeptide.

Conclusions
The broad-spectrum lipopeptide showed promising antifungal activity against dermatophytes and may be considered for nano-emulsion formulation and tested for topical application in a mice model.
P092 Otilonium bromide is a potent antifungal agent against fluconazole- and flucytosine-resistant Cryptococcus neoformans strains
Authors: Zhen Cheng , Lu Hui , Jiang Yuan-ying , Yang Feng 
Published: September 2022

Abstract

Poster session 1, September 21, 2022, 12:30 PM - 1:30 PM

Objectives
Cryptococcus neoformans is a worldwide threat causing global pulmonary and systemic infections in humans. However, only three drugs are available for the treatment of cryptococcosis: fluconazole, amphotericin B, and flucytosine. Drug repurposing, the process of using a drug for an indication different from the initial indication, is an emerging approach in the discovery of new antifungal drugs. We've set out to identify repurposable drugs for cryptococossis.

Methods
We screened the United States FDA-approved drugs for antifungal activity against C. albicans lab strain SC5314. We tested the antifungal profile against major human fungal pathogens including C. neoformans. In order to uncover the mechanism of action against C. neoformans, lab strain H99 was used to evolve drug resistant adaptors. Next-generation sequencing technology was used to investigate the genome change of drug resistant adaptors. SNP calling was also performed to identify possible mutations causing drug resistance.

Results
Here we found that otilonium bromide (OTB), which is extensively used to treat patients affected by the irritable bowel syndrome, had broad-spectrum antifungal activity. OTB was active against fluconaozle-resistant and flucotysine-resistant C. neoformans strains. Furthermore, we found resistance to OTB was mostly due to duplication of chromosome 6. Further work will be on the identification of the Candidate gene on chromosome 6 which is required OTB resistance.

Conclusion
This study highlights the potential application of OTB as a new antifungal drug against C. neoformans strains susceptible or resistant to commonly used antifungal drugs.
P093 A novel GAL4-like transcriptional regulator modulate the azoles sensitivity of Exophiala dermatitidis
Authors: Zhu Yutang , Zhang Jie , Li Xiaotong , Sun Yi , Gao Lujuan 
Published: September 2022

Abstract

Poster session 1, September 21, 2022, 12:30 PM - 1:30 PM

Objectives
Exophiala dermatitidis is an emerging opportunistic pathogen among the ever-increasing numbers of immunocompromised hosts. Understanding its transmission mechanism, pathogenicity, and potential azole-resistant mechanism is important for better understanding of this fatal pathogen and benefits future diagnosis, and treatment.
By using a PCR-based fragment fusion approach, we applied phosphotransferase (HPH) resistance gene which amplified from pAN7-1 as a selective gene for generating replacement cassettes. Combined with electroporation transformation and screening on selection medium containing 50 μg/ml HMB, we obtained the gene knockout strain Δ GAL4, a novel GAL4-like transcriptional factor (XP_009154426.1) encoding gene (GeneID:20 306 783) accurately deleted.

Methods
Growth curve determination: Take 200 μL of 2 × 10⁴ CFU/ml bacterial suspension and 200 μL liquid sabourand medium in EP tubes. Shaking at 200 rpm 37°C for 0 h, 1 h, 2 h, 3 h, 4 h, 8 h, 12 h, 16 h, 18 h, 24 h, 30 h, 36 h, 48 h. Then add 200 μL cultured bacterial suspension to a 96-well plate and repeat three times. Meanwhile set up blank medium control wells. By using an enzyme marker to measure the absorbance at 504 nm, analyzing with SPSS, and then graphing with GraphPad software.
Morphological observations: Activated strains on Sabourand microcultures and incubated at 37 °C for 3d. Observe morphology by using fluorescent microscopy to compare the different strains of wild and knockout strains, and using lactic acid cotton phenol blue/fluorescent dye filming observe the morphology of conidia.
E-test drug sensitivity method: Spread fresh bacterial suspension of ΔGAL4 and ATCC34100 containing 1 × 10⁶CFU/ml spores on RPMI 1640 agar plates with sterile cotton swabs, and place drug sensitivity test strips [itraconazole (Itraconazole Liofilchem, ICZ 92148), posaconazole (Posaconazole Liofilchem (PCZ 92152), voriconazole (Mérieux, VRC 532800)] in the center plate, after sealing with a parafilm, incubating in 37°C for 36-48 h, and read the values at the tangent of the drug susceptibility strip and the antibacterial circle.

Results
The results revealed that the ΔGAL4 showed almost no difference in growth and morphology compared with wild strain, but its sensitivity to azoles was altered. We applied E-test to evaluate the MIC changes of azoles. The MIC of itraconazole, voriconazole, and posaconazole against ΔGAL4 was 0.5 μg/ml, 0.125 μg/ml, and 0.25 μg/ml; against ATCC34100 was 2 μg/ml, 0.19 μg/ml, and 0.38 μg/ml, respectively. △GAL4 MIC suggests that ∆GAL4 showed more sensitivity to azoles compared to its wild-type strain.

Conclusion
The present results suggested that a novel GAL4-like transcriptional factor may play an important role in azole sensitivity of E. dermatitidis, which may be through modulating efflux pumps or Cyp51A. Further investigation of this GAL4 related modulating pathway is needed.