bluuebunny/crossref_metadata_2025_split huggingface dataset:
P446 Molecular detection of fungal agents responsible for COVID-19-associated mycosis directly from tissue specimens
Authors: Singh Vikramjeet , Das Anupam , Agarwal Ashish Chandra , Gupta Nikhil , Sen Manodeep , Agarwal Jyotsna 
Published: September 2022

Abstract

Poster session 3, September 23, 2022, 12:30 PM - 1:30 PM

Objective
Due to the ongoing COVID-19 pandemic a new group of patients at risk emerged with COVID-19-associated mucormycosis (CAM) and other fungal infections. Molecular studies, evaluating the prevalence of CAM and other fungal infections are lacking. To assess CAM prevalence in a super-specialty healthcare hospital in North India, we applied direct microscopy, fungal culture, and qualitative real-time-PCR targeting Mucorales-specific fragments on tissue specimens of critically ill COVID-19 patients.

Methods
This was a hospital-based prospective study during second-wave of COVID-19 in India. All clinically suspected CAM patients with a history of COVID-19 were included in the study from March 2021 to June 2021 where tissue or biopsy specimens were collected under aseptic conditions. Conventional identification methods were performed for all isolates, speciation was done by MALDI-TOF, and comparative detection by RTPCR was also done.

Results
In the present study, among 67 samples received in the laboratory from clinically suspected CAM patients, 32 samples showed positive growth using the conventional method of identification. Rhizopus arrhizus was the commonest fungal isolate obtained followed by Aspergillus flavus from tissue samples. Use of molecular and automated machines helped in the early identification of these species 24-48 h less than the conventional methods. Polyfungal isolates are also reported from two tissue samples of patients in the post-COVID discharge stage. Almost 90% of patients with CAM and other fungal etiology agreed to steroid intake and diabetes condition during COVID-19 infection.

Conclusion
Considering the ever-evolving strains and variants of COVID-19, it is important to have a high index of suspicion for fungal coinfection in patients with COVID-19 presenting with comorbidities. Further, they should undergo immediate molecular studies with an emphasis on the requirement of medical or surgical intervention if the result comes positive. There is a need to stress on the judicious use of steroids to avoid flaring up of the fungal infection.
P459 The Aspergillus lateral flow assay for the diagnosis of chronic pulmonary aspergillosis
Authors: Peng Jie , Yan Jun , Zhang Yuan , Yao Wang , Wang He , Su Yan , Zhou Zeqi 
Published: September 2022

Abstract

Poster session 3, September 23, 2022, 12:30 PM - 1:30 PM

Objectives
Chronic pulmonary aspergillosis (CPA) is an uncommon and problematic pulmonary disease, complicating many other respiratory disorders. Measurement of Aspergillus-specific IgG antibodies had a higher sensitivity than either IgM, IgA, or IgE and it should therefore be considered the most appropriate test for screening CPA. Aspergillus-specific IgG antibody levels have been successfully used to monitor the response of CPA to medical therapy. Recently, a novel rapid test for Aspergillus-specific IgG antibody from Dynamiker Biotechnology (Tianjin) Co., Ltd. was released as a screening test of CPA. It is a fluorescent immunochromatographic cassette test using a monoclonal antibody against Aspergillus-specific IgG antibody and Europium nanoparticles (Eu NP) (Fig. 1). It is a semi-quantitative test which was easy to operate, rapid with portable detection devices, and can be widely accepted by clinical and primary medical (Fig. 2). We collect clinical samples to verify the detection performance of the Dynamiker Quic™Aspergillus-specific IgG antibody (LFA) test.

Methods
In total, 102 patients were included and 42 patients were diagnosed with CPA. The pathogen was identified from sputum, BALF culture, lung resection surgery, bronchos copy biopsy, percutaneous lung biopsy and BALF GM assay.

Results
The sensitivity, specificity, PLR, NLP and Kappa index of the IgG antibody test were 85.6%,94.4%,14.0%,0.23% and 0.72, respectively.

Conclusions
The current work indicates that the Dynamiker Quic™ Aspergillus specific IgG antibody (LFA) test shows a promising application in the diagnosis of CPA. The results are accurate and reliable, and it could be used as an aid for the early rapid screening test of CPA.
P473 Genomic epidemiology of antifungal-resistant Candida auris in Colombia
Authors: Escandon Patricia , Misas Elizabeth , Gade Lalitha , Caceres Diego , Hurst Steve , Litvintseva Anastasia , Duarte Carolina , Chow Nancy A. 
Published: September 2022

Abstract

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

Introduction
Candida auris is a public health threat. Five major clades of C. auris have been identified (Clades I–V). In Colombia, C. auris infections were first reported in 2016 with ongoing transmission reported from multiple cities. Here, we describe C. auris genomic epidemiology in Colombia detailing cases from 2016–2021.

Methods
A total of 99 isolates from C. auris cases were collected between June 2016 to January 2021 in Colombia, representing 11 geographic locations. Species confirmation, antifungal susceptibility testing, and whole-genome sequencing (WGS) were performed. In all, 37 genomic sequences generated previously from isolates from C. auris cases in Colombia, Venezuela, Panama, Israel, and United States were also analyzed MycoSNP workflow was used to assess sequence quality, map reads to the reference, and identify single-nucleotide polymorphisms (SNPs). Pairwise distances and a neighbor-joining tree were generated. IQtree was used to generate a maximum-likelihood tree with bootstrap values.

Results
Phylogenetic analysis identified 1 493 SNP positions. Isolates from Colombia clustered to Clade IV and predominately grouped by country except for 16 fluconazole-resistant isolates from Bogota, Colombia that grouped with five isolates from Venezuela. In this cluster, 20 (95%) were resistant to fluconazole and 5 (24%) were resistant to fluconazole and the echinocandin micafungin. Remaining isolates from Bogota did not group in this cluster and were susceptible to fluconazole and micafungin.
A total of 98 isolates from Colombia clustered together. Within this Colombian cluster, there were two subgroups that had bootstrap support of 100% and were separated by 13 SNPs. The first subgroup was a cluster that contained 18 isolates from the north coast; 17 (94%) isolates were resistant to amphotericin B. A second subgroup consisted of 26 isolates from Cesar and Norte de Santander, and 22 (84%) isolates were resistant to fluconazole.

Conclusions
Based on the phylogenetic reconstruction, C. auris in Colombia continues to be of Clade IV. Amphotericin B-resistant isolates were predominately from the north coast, fluconazole-resistant isolates were from a wider geographic area in Colombia, and echinocandin-resistant isolates were from Bogota. Within the Colombian cluster comprising two subgroups, we observed high genetic relatedness between isolates from different geographic locations suggesting transmission among cities.
P474 The value of PCR-based azole resistance detection in invasive aspergillosis: A prospective multicenter study
Authors: Huygens Sammy , Dunbar Albert , Buijl Jochem , Maertens Johan , Verweij Paul , Klaassen Corne , Lagrou Katrien , Van Dijk Karin , Mercier Toine , Schauwvlieghe Alexander , Rijnders Bart 
Published: September 2022

Abstract

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

Objectives
Prompt detection of azole-resistant Aspergillus fumigatus will result in the timely start of active treatment and may improve the survival of invasive aspergillosis (IA). The use of a multiplex polymerase chain reaction (PCR) targeting Aspergillus species and fumigatus DNA as well as the two most prevalent azole resistance- associated mutations (RAMs) in the cyp51A gen (TR34/L98H and TR46/Y121F/T289A) could shorten the time to detect azole-resistant IA.

Methods
In a prospective study in 12 Dutch and Belgian centers, we evaluated the clinical value of the multiplex AsperGenius®PCR in hematology patients with a pulmonary infiltrate undergoing bronchoalveolar lavage (BALf) sampling. The primary endpoint was antifungal treatment failure in the 6 weeks after antifungal treatment initiation in the patients in which azole-resistant IA was detected. Treatment failure was defined as death or a switch to an antifungal agent from another class after at least 5 days of first-line therapy. Patients with a mixed azole-susceptible/resistant infection were excluded from this analysis to ascertain that the infection was indeed caused by the resistant strain.

Results
Of 323 patients enrolled, sufficient BALf for PCR testing remained in 299. Probable fungal disease was diagnosed in 95 (34%), Aspergillus cultured in 24 (8%), Aspergillus DNA detected in 118 (39%), and A. fumigatus DNA in 88 (29%) patients. The resistance PCR was conclusive in 54/88 (61%) and RAMs were detected in 8 (15%), Table 1. All 8 had probable IA but 2 had a mixed infection and were excluded. In the 6 remaining patients, treatment failure was observed in one. Compared with the GM negative patients and despite antifungal therapy, a positive GM test was associated with a 13% higher 6-week overall mortality (P = .01), Table 2. Surprisingly, the 6-week mortality in the 65 patients who had a positive Aspergillus PCR but a negative GM and culture was not increased compared to those with a negative PCR (PCR + 14% vs. PCR- 16% mortality, P = .68).

Conclusions
In patients with an underlying hematological disease and a pulmonary infiltrate, the detection of Aspergillus DNA by PCR on BALf was not associated with increased mortality. The exact place of the Aspergillus PCR in the EORTC-MSGERC invasive fungal infection criteria is therefore uncertain. In 15% of the patients in whom A. fumigatus DNA was present, azole RAMs were detected by PCR. In only 1/6 probable cases of IA with RAMs detected, antifungal treatment failure was observed. Basing the choice of antifungal therapy on the result of a cyp51a resistance PCR may help to reduce the impact of azole resistance on mortality.
P475 Bee health and the antifungal activity of honey
Authors: Fernandes Kenya , Stanfield Bridie , Susantio Daniel , Carter Dee 
Published: September 2022

Abstract

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

Objectives
Various components have been identified which contribute to the antimicrobial properties of honey, many of which are secreted by bees into the honey as it is being processed. Here we investigate the relationship between bee and hive health and the antifungal properties of honey.

Methods
Samples of honey were collected from hives that were either healthy or experiencing some kind of distress as assessed by beekeepers. Healthy hives displayed strong population numbers and good brood patterns while distressed hives exhibited warning signs such as low population numbers, patchy brood patterns, evidence of chalkbrood fungal disease, or small hive beetle infestation. Honey samples were tested for antifungal activity against yeast and mold species via broth microdilution, tested for hydrogen peroxide levels via colorimetric assay, and spread on agar plates to assess the abundance and diversity of microbes present in the raw honey.

Results
Honey samples were effective against the yeast Cryptococcus deuterogattii and the dermatophyte mold Trichophyton interdigitale but ineffective against the yeast Candida dubliniensis and the mold Aspergillus flavus. The hydrogen peroxide levels of the honeys were variable and did not always align with activity. Less microbes in number and abundance were present on agar plates grown from healthy hive honey compared to distressed hive honey.

Conclusion
These results indicate that bee health may play a role in contributing to the antifungal properties of honey and that promoting strong and healthy hives is beneficial.
P476 Monitoring antifungal resistance in a global collection of Candida spp. surveillance isolates, including C. auris—analysis of resistance in antifungals (ARIA) 2020 study
Authors: Kothari Nimmi , Hawser Stephen , Olari Sara , Sartori Silvia , Mathur Tarun 
Published: September 2022

Abstract

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

Objectives
Analysis of resistance in antifungals (ARIA) is a recent longitudinal global surveillance initiative collecting yeast and fungal isolates from hospitals worldwide designed to determine susceptibility profiles and monitor the resistance trends among antifungal agents. ARIA reports the susceptibility patterns of data concerning echinocandins, second-generation triazoles, and fluconazole against clinical Candida spp., and filamentous fungal isolates from worldwide sources.

Methods
Candida spp. isolates (n = 662) were collected from hospitals worldwide during 2020 from 13 different sites—Argentina (n = 1), Australia (n = 2), Germany (n = 1), India (n = 2), Italy (n = 1), Panama (n = 1), Spain (n = 1), Turkey (n = 1), United Kingdom (n = 1), and United States (n = 2). These isolates were shipped to a central laboratory at IHMA Europe, Switzerland, and re-identified by MALDI-TOF or molecular methods. The MICs were performed by broth microdilution method in line with CLSI susceptibility testing standards—CLSI M27-A4 and M38-A2—methodologies and percentage susceptibility (%S) were calculated. Antifungals tested were amphotericin B (AMB), anidulafungin (ANID), fluconazole (FLU), isavuconazole (ISA), caspofungin (CASP), micafungin (MIC), posaconazole (POS), and voriconazole (VOR).

Results
Table 1: Summary MIC and susceptibility data of Candida spp. isolates for all countries combined

Conclusions
The data from the ARIA 2020 study indicate that overall antifungal resistance is low among the Candida spp. isolates except for C. glabrata and C. krusei where resistance to one or more antifungal agents was observed. However, there was no significant difference in susceptibility pattern was observed when susceptibility data of C. glabrata and C. krusei from different continents were compared. The emergence of resistance was evident among C. auris isolates as they have shown reduced susceptibility to azoles in this study.
Antifungal resistance surveillance and investigation into resistance mechanisms are of paramount importance. The ongoing ARIA surveillance study will provide resources to monitor antifungal resistance trends, provide key information to caregivers and provide essential information with respect to the development of novel antifungal agents.
P497 Candida DNA extraction from positive blood culture bottles
Authors: Balakumar Uthra , Subramanian Anitha , Kindo Anupma Jyoti 
Published: September 2022

Abstract

Poster session 3, September 23, 2022, 12:30 PM - 1:30 PM

Background
Molecular technique of detecting candida from blood is a quick method to identify candidemia. A major limitation of the molecular method is the difficulty associated with breaking fungal cell wall, the DNA extraction step still requires more than half of a working day.
The successful extraction of DNA involves effective disruption of cells, denaturation of protein, and nucleoprotein complexes, and inactivation of nucleases such as DNase. The extracted DNA should have low contamination of proteins.

Objectives
To perform DNA extraction using phenol-chloroform method.
2. To speciate candida using PCR-RFLP.

Methods
Candida DNA is isolated using phenol-chloroform method. The PCR products of different candida species obtained directly from blood samples were subjected to restriction enzyme (MspI).

Results
DNA was extracted from 50 samples, out of which 40 samples were from known positive blood culture bottles and the remaining 10 were known negative for candida. Of the 40 known positive blood culture bottles, 34 produced bands of various molecular weights and all 10 negative cultures did not show bands. Representative amplicon is shown in the gel picture (Figure 1).
Out of 40 candida-positive blood culture samples, 7 species have been identified using PCR- RFLP method.

Conclusion
The turn-around time can be highly reduced by extracting DNA from positive blood culture bottles and then performing PCR and RFLP for speciation compared with the conventional method.
S1.5d Spectrum of etiological agents of mycotic keratitis: A n 11- year review
Authors: Thomas Philip Aloysius , Kaliamurthy Jayaraman 
Published: September 2022

Abstract

S1.5 Mycotic Keratitis, September 21, 2022, 11:00 AM - 12:30 PM

Background
To assess the spectrum of etiological agents of mycotic keratitis over an 11-year-period at a tertiary eye care center in southern India.

Methods
A retrospective review was made of microbiological data relating to corneal scrapings performed over a period of 11 years (January 2011-December 2021) on 1200 individuals who presented with suspected microbial keratitis. Each individual underwent corneal scraping and the scraped materials were subjected to meticulous microbiological analysis that included direct microscopy (Gram-stain and lactophenol cotton blue wet mount) and culture on multiple solid and liquid culture media.

Results
A total of 404 fungal isolates were recovered from the corneal scrapings of 1200 patients with suspected microbial keratitis. Of the 404 fungal isolates, Fusarium spp (133) were the predominant isolates, followed by Aspergillus flavus (104), Curvularia spp (24), Aspergillus fumigatus (17), Bipolaris sp (7), Alternaria sp (2), Colletrotrichum sp (2) Cylindrocarpon lichenicola (2) Exserohilum sp (1) and Drechslera sp (1); there were also 111 filamentous fungus isolates that defied identification in spite of various efforts made to induce sporulation. Of the 404 culture-proven cases of mycotic keratitis, 381 patients confirmed ocular trauma while engaged in agricultural activity.

Conclusion
Fusarium spp., followed by Aspergillus spp., were the most common organisms found in mycotic keratitis patients in this specific geographical area. Additional efforts are required to spread awareness among villagers about the dangers of not promptly treating mycotic and other forms of microbial keratitis so that blindness and visual disability caused by corneal scarring in rural areas can be reduced.
S10.3d Yeast Microbiome in patients with inflammatory bowel disease (IBD)
Authors: Vaezi Afsane , Fakhim Hamed , Jabbari Mehrdad , Nasri Elahe , Azar Mohammad Reza Hosseini , Diba Kambiz , Badali Hamid 
Published: September 2022

Abstract

S10.3 The mycobiome characterization: future perspectives or just a trend?, September 24, 2022, 10:30 AM - 12:00 PM

Objective
The intestinal microbiota plays major roles in host-protective functions and inflammatory bowel diseases (IBDs). Although some data suggest a role of the yeast microbiota in IBD pathogenesis, the available data are rare. The aim of this study was to evaluate the fecal yeast microbiota in patients with IBD.

Methods
Fungal isolates of the fecal microbiota of 120 patients with IBD and 30 healthy control individuals were determined using conventional and molecular (ITS sequencing and RFLP) methods.

Results
The mean age of the patients was 37.4 ± 12.2 years (range 15-72 years). Of 120 patients included, 67 had ulcerative colitis (55.8%) and 53 had Crohn's disease (44.2%). Ulcerative colitis was more prevalent in women than Crohn's disease (61.7% vs 50.2%, P &lt;.005). The median time between the onset of symptoms and diagnosis was 9.58 ± 8.56 years. In total, 73 colonies of Candida spp. were isolated from 60 patients with IBD. The most common identified species of Candida were C. albicans (42.4%), C. glabrata (20.5%), and C. krusei (6.8%). The incidence of non-albicans Candida species (57.6%) was higher than C. albicans (42.4%). We observed an increased proportion of C. albicans compared with healthy individuals (28.5% vs 21.6%). The frequencies of C. glabrata were significantly higher among IBD patients rather than the control group (20.5% vs 9.8%).

Conclusion
Candida albicans were found to be increased in abundance in the IBD samples. These data emphasize the potential importance of yeast microbiota signatures as biomarkers. Moreover, we unravel here disease-specific-Candida species network alterations in IBD, suggesting that, Candida species might play a role in IBD pathogenesis.
S10.4b Pathophysiology, diagnosis, and management of chronic pulmonary Aspergillosis
Authors: Izumikawa Koichi 
Published: September 2022

Abstract

S10.4 Emerging antifungal resistant fungi, September 24, 2022, 10:30 AM – 12:00 PM

Chronic pulmonary Aspergillosis (CPA) is a complex disease that is difficult to diagnose and resistant to treatment. Many cases are missed and have unfortunate outcomes in clinical settings. Although CPA has long been classified into several types based on pathological findings, it is not always possible to make a pathological diagnosis in all cases, so a clinical diagnosis is often made. In addition, many have an underlying respiratory disease and complications by infections caused by other microorganisms, making treatment further difficult. The diagnosis of CPA requires clinical symptoms and findings, radiological, serological, pathological, and microbiological approaches, and standardization of these diagnostic criteria is challenging. The mainstay of treatment is outpatient oral therapy with azoles, and indications and therapeutic evidence for echinocandins, and polyenes are limited. The development of novel antifungals with different mechanisms of action from conventional agents is also awaited. Azole-resistance of Aspergillus caused by long-term treatment is also an issue. It has been reported that resistant strains have been found in CPA patients treated with long-term azole therapy, mutations in the cyp51A and others have also been identified and longer exposure to azoles are correlated with higher MIC values of them. This presentation will focus on epidemiology, pathogenesis, diagnosis, and management including drug resistance in CPA patients.
S10.4d A unique clinical appearance of Candida auris infection in Japan
Authors: Umeyama Takashi , Abe Masahiro , Shinohara Takayuki , Miyazaki Taiga , Miyazaki Yoshitsugu 
Published: September 2022

Abstract

S10.4 Emerging antifungal resistant fungi, September 24, 2022, 10:30 AM - 12:00 PM
It has only been 15 years since Candida auris was reported isolated from the ear canal of a 70-year-old Japanese woman in Tokyo, and no record of an isolate corresponding to this species has been found prior to 1996. It is a high public health priority concern in several regions of the world. This is because the fungus is multidrug-resistant and can acquire resistance to all three major groups of current antifungal drugs (azoles, echinocandins, and amphotericin B). Outbreaks in healthcare facilities are also a concern. The main reasons for this are as follows: unlike other Candida spp. that primarily inhabit the digestive and urinary systems, C. auris readily colonizes patient skin and can survive for several weeks on dry, non-living surfaces, contributing to infections and outbreaks in healthcare facilities.

In Japan, C. auris was first identified in 2009 in a discharge from the ear canal of a patient admitted to a Japanese hospital, and since then, all isolates have come from the ear canal, with only a few reported strains. For reasons unknown, as of 2022, C. auris has not been reported as a cause of invasive disease in Japan, and no nosocomial infections have occurred. Whole genome analysis suggests that all Japanese isolates belong to Clade II, affecting drug resistance and clinical characteristics.

In this symposium, I will present the current status of C. auris infection in Japan, the first country where C. auris infection originated, together with its unique clinical features and molecular epidemiological analysis.