bluuebunny/crossref_metadata_2025_split huggingface dataset:
P452 Estimation of the detection limit of extracted Candida DNA from spiked EDTA blood sample
Authors: Subramanian Anitha , Balakumar Uthra , Kindo Anupma Jyoti 
Published: September 2022

Abstract

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

Background
Candidemia caused by genus Candida, are opportunistic pathogens with life-threatening infection in immunocompromised individuals with a mortality rate being 40%–54% in patients. Blood culture is not a reliable and rapid method for the diagnosis of candidemia as its sensitivity is as low as 50%. Molecular diagnosis is an alternative to conventional culture for early detection of candidemia. An appropriate DNA isolation technique is needed to obtain high purity DNA from blood specimens and improves the sensitivity of the polymerase chain reaction (PCR). In this study, two different methods were used to extract Candida DNA from spiked EDTA blood specimens, the In-house phenol-chloroform and potassium acetate method.

Objectives
DNA extraction from the spiked blood samples
Quantification of the Candida DNA extracted from the spiked sample
Comparison of the extracted DNA by phenol-chloroform and potassium acetate method

Methods
A total of 5 ml of EDTA blood samples from healthy volunteers were spiked with 104-108 Candida albicans cells (ATCC 90028) to determine the detection limit of our extraction method. DNA was extracted from whole blood using phenol-chloroform and the potassium acetate methods which involves pretreatment with erythrocytes, leukocytes and fungal lysis buffer. DNA from C. albicans were amplified using ITS1 and ITS4 based primers. PCR products were visualized by agarose gel electrophoresis.

Results
The spiked Candida cells taken were from 104-108 yeast cells. The total extracted DNA by phenol chloroform extraction method ranged from 18.29 to 51.64 ng/μl respectively. Whereas the total extracted DNA from potassium acetate extraction method ranged from 15.58 to 32.30 ng/μl respectively. The presence of a 535 base pair (bp) product was considered positive (Figs. 1 and 2). The lowest detection limit of PCR has been determined as 106-108 Candida cells for our spiked samples.

Conclusion
In our study, till date, the extracted quantity of DNA was found to be higher by phenol-chloroform method as compared to the potassium acetate method. Greater numbers and processing are required to obtain conclusive data.
P466 Isolation of Cryptococcus neoformans and other yeast from pigeon droppings in Khartoum state, Sudan
Authors: Abdalla Wisal 
Published: September 2022

Abstract

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

Objectives
The aim of this study was to investigate the presence of Cryptococcus neoformans and other yeast species in pigeon droppings in Khartoum state, Sudan.

Methods
Sample collection A total of 120 samples were collected from pigeon droppings from Khartoum state. Pigeon droppings were collected using sterile wooden spatulas and placed directly in clean plastic bags.
Processing of sample
Approximately 2 g of pigeon droppings were added to 10 ml of sterile saline. The samples were allowed to stand for 20 min with frequent vortex.
Isolation and Identification of yeast isolates
A loopful of supernatant fluid from prepared samples was taken and streaked onto Sabouraud's dextrose agar media supplemented with 0.05 mg/ml chloramphenicol incubated at 37°C for 1-2 days. All yeast isolates were identified by direct microscopic examination using the lacto-phenol-cotton-blue stain India ink stain and Gram's stain. Further confirmatory tests were conducted using Corn Meal Agar (CMA) with tween 80, germ tube production test using horse serum, and urease test. Confirmation of identification was done using API 20C AUX and API ID 32C AUX (bioMérieux®, Madrid, Spain).

Results
Cryptococcus neoformans is the most common species isolated from pigeon droppings as shown in Table 1. Different Candida species have been isolated Figure 1.

Conclusion
The present study concluded that there is a potential role of pigeons as a reservoir for C. neoformans and other zoonotic yeasts in the environment that can affect humans and animals.
P478 Identification of clinical Candida isolates using maldi-tof-ms and their antifungal susceptibility profile—a study from tertiary care pediatric hospital in noida
Authors: Perween Naz , Nandwani Sumi , Verma Nikhil , Rai Sumit , Prakash M.R. Shiva , Saxena Krati , Dwivedi Niharika 
Published: September 2022

Abstract

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

Introduction

Objectives

Material and Methods
The study was conducted in Department of Microbiology during the period of August 2018 to August 2021.

Results

Conclusion
Rapid identification of Candida species using Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) as a reliable and rapid technique for Candida species identification as candida is an important cause of neonatal sepsis and sepsis related mortality.

Sources
P484 Expanding VGVI—evidence for distinct Cryptococcus gattii (decagattii) endemic to the American Southwest
Authors: Engelthaler David , Monroy-Nieto Juan , Sykes DVM Jane , Komatsu Ken , Meyer Wieland 
Published: September 2022

Abstract

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

Objectives
We aimed to understand the nature of autochthonous Cryptococcus gattii clinical and veterinary cases identified in Arizona, a state in the American Southwest, a locale well outside of the known endemic regions.

Methods
Whole-genome sequencing and phylogenomic comparative analyses were conducted on four unrelated isolates collected from recent cases along with other relevant C. gattii genomes.

Results
Phylogenomic analysis grouped the Arizona genomes with a previously known set of Mexican isolate genomes, labelled as VGVI or C. decagattii. These genomes are clearly delineated from the nearest major molecular type (VGIII), but show no recombination with other molecular types or species of C. gattii.
See Figures below.

Conclusion
These findings expand VGVI into a definitive clade and establish this molecular type as a clinically important and distinct population. These findings also expand the known Cryptococcus ecological range into a previously unrecognized endemic area, typified by extreme heat and aridity.
P488 Successful treatment of breakthrough invasive aspergillosis in an immunocompetent individual based on therapeutic drug monitoring: A case report
Authors: Yin Xin , Zong Zhiyong , Liu Yanbin 
Published: September 2022

Abstract

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

Background
Invasive aspergillosis (IA) is an opportunistic fungal infection in immunocompromised patients with high mortality. Aspergillus flavus is the second pathogen of IA. Breakthrough IA was defined as any IA occurring during exposure to an antifungal drug.

Case presentation
A 22-year-old female college student was admitted severely unwell with dizziness and left limb weakness. She was healthy previously and did not take any medication. Magnetic resonance imaging showed a right intracranial space-occupying lesion. The postoperative pathological and morphological examinations suggested Aspergillus flavus. The anti-fungal medication, voriconazole, was administered immediately. Unfortunately, her condition deteriorated, and she experienced coma after about 1 month of antifungal treatment. The emergency craniotomy revealed a large amount of pus and the culture of pus confirmed Aspergillus flavus. Antifungal regimen was developed by infectious disease specialists, and drug concentration was monitored continuously. This patient received antifungal treatment for 2 years. No recurrence was observed after 6 months of antifungal drug withdrawal, and she can take care of herself.
See Figures below.

Conclusion
Breakthrough IA occurs in patients who lack high risk factors, making diagnosis more difficult and leading to a higher risk of mortality. Therapeutic drug monitoring is crucial for therapeutic success. Meanwhile, multidisciplinary therapeutics can improve the survival rate.
P490 Fatal secondary fungemia due to Trichosporon asahii onychomycosis in a diabetic patient
Authors: Taj-Aldeen Saad , Francisco Elaine , Al Maslamani Muna , Theelen Bart , Vu Duong , Boekhout Teun , Hagen Ferry 
Published: September 2022

Abstract

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

Objectives
We describe a fatal case of trichosporonosis caused by Trichosporon asahii. The aim was to molecularly characterize the T. asahii strains from blood and foot tissue samples to investigate their genetic relatedness.

Case
An 85-year-old morbidly obese female with a prior cerebrovascular accident, hypertension, and diabetes mellitus was admitted to a peripheral hospital with type II respiratory failure, metabolic acidosis, and chronic anemia. Three weeks post-hospitalization the patient remained febrile, physical examination showed that the patient had paronychia, nail pigmentation, subungual onychomycosis, and a diabetic foot ulcer. Blood culture, as well as nail and ulcer samples, became positive for Trichosporon yeasts.

Methods
Trichosporon yeasts were subjected to molecular identification by sequencing the intergenic spacer (IGS1) region. Minimal inhibitory concentrations (MICs) were determined by the EUCAST microdilution method. Long-read nanopore sequencing was performed for the three clinical strains, the type-strain of T. asahii (CBS2479), and two IGS-genotype 7 strains (CBS2936, CBS7632) using the native barcoding kit v12 (SQK-LSK112; Oxford Nanopore Technologies). Raw data were basecalled with Guppy v6, Flye v2.9 was used to de novo assemble the genome of CBS2479, this was used as a reference for variant calling using the genomic reads of all strains.

Results
The three clinical strains were found to belong to the rare IGS-genotype 7 and had similar MICs for amphotericin B (4 µg/ml), 5-fluorocytosine and fluconazole (2 µg/ml), voriconazole (≤ 0.015 µg/ml), posaconazole (0.0625 µg/ml), itraconazole (0.125 µg/ml), caspofungin (8 µg/ml), anidulafungin and micafungin (&gt; 16 µg/ml). Strains from the ulcer and nail were genetically very closely related with 10 836 SNPs differences, the blood-derived strain differed more from these two strains with 94 729 SNPs and 94 913 SNPs, respectively. Strains CBS2936 and CBS7632 were only distantly related to CBS2479 with 328 221, and 436 060 SNPs, respectively.

Conclusion
The T. Asahii IGS-genotype 7 strain causing fatal fungemia was genetically nearly identical to that obtained from nail and foot ulcers, making it highly likely that this was the port d'entrée for T. asahii.
P492 Invasive fungal infection during COVID era at tertiary care hospital
Authors: Seema Kumari , Kumar Abhay , Kumar Manoj , Sharma Ashok , Boipai Manju 
Published: September 2022

Abstract

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

Introduction
Invasive fungal infection is a leading cause of morbidity and mortality in COVID patients as a secondary infection. Invasive fungal infections are often rapidly life-threatening and require specialist consultation for prompt diagnosis and therapy. The reporting of these invasive fungal infections (Mucormycosis, Aspergillosis, and candidiasis) is less than its occurrence.

Objective
This study was undertaken to diagnose invasive fungal infections in COVID patients during the first, second, and third COVID-19 pandemic waves by conventional methods.

Materials and Methods
From March 2020 to December 2020 was considered the first wave pandemic, March 2021 to May 2021 was the second wave pandemic and December 2021 to February 2022 was considered the third wave pandemic era for this region. A total of 42, 168, and 87 samples were collected from patients clinically suspected to have invasive fungal infections during three pandemic waves from Covid indoor patients of RIMS, Ranchi. Among a total 24 were tissue samples, 13 BAL and 260 were nasal swabs/scrapes collected from suspected patients. KOH screening of all samples was done followed by culture on SDA media.

Results
A total of 83/297 (27.94%) cases were positive for KOH screening. In all, 91/297 (30.64%) samples showed culture positivity. A majority of growth was for Mucorales (48.2%), Aspergillus species (24.1%), Candida species (15%), and mixed growth (12.7%). Among Mucorales, Rhizopus was identified in 64% followed by Mucor (28%), Absidia (5%), and Rhizomucor (3%). Among Aspergillus species, majority were A. flavus (48%) followed by A. niger (37%), and A. fumigatus (15%). Maximum positivity for invasive fungal infections was observed during the second wave pandemic (62%) followed by the third wave (27%), and the first wave (11%) respectively.

Discussion
Maximum cases were observed during and after the second wave COVID pandemic era due to excessive use of steroids during COVID treatment and cases were decreased during the third wave as compared with second wave timeframe due to the use of COVID vaccine, and many asymptomatic cases and home quarantine policy.

Conclusion
Early diagnosis and treatment of invasive fungal infections with antifungal therapy and surgical debridement are necessary to reduce mortality and end-organ damage.
P504 Screening of Belgian bats and hibernacula for the description of related fungal microbiomes and the detection of Pseudogymnoascus destructans
Authors: Debergh Hanne , Becker Pierre , Van den Eynde Claudia , Baert Frederik , D'hooge Elizabet , De Pauw Robby , Normand Anne-Cécile , Piarroux Renaud , Stubbe Dirk 
Published: September 2022

Abstract

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

Bats can be affected by fungal pathogens such as Pseudogymnoascus destructans, the causative agent of the white-nose syndrome. Their body surface can also be colonized by fungal commensals or carry transient fungal species and participate in their dispersal. The present study aimed to assess the presence of P. destructans in Northern Belgium, to describe the skin mycobiome of active bats during summer and autumn, and to analyze possible differences in fungal diversity among bat species, sampling sites, and seasons. In total, 114 bat specimens belonging to seven species were sampled from various localities. Culture-based methods revealed an important mycological diversity with 209 different taxa. Overall, a mean of 3.7 taxa per bat was recorded but significant differences were observed between sampling sites and seasons with a higher diversity in autumn as compared to summer. The mycobiomes were dominated by cosmopolitan and plant-associated species, in particular from the genera Cladosporium, Penicillium, and Aspergillus. Other species known to be related to bats or their environment, like Apiotrichum otae, were also retrieved. Although P. destructans was not detected, the sampling of the hibernacula indicated that they can be inhabited by diverse fungal species including a yet undescribed Pseudogymnoascus species, distinct from P. destructans, namely P. cavicola, sp. nov.
S1.3d The human pathobiont Malassezia furfur secreted protease MfSAP1 regulates cell dispersal and exacerbates skin inflammation
Authors: Goh Joleen , Ruchti Fiorella , Poh Si En , Koh Winston L.C. , Tan Kiat Yi , Lim Yan Ting , Thng Steven T.G. , Sobota Radoslaw M. , Hoon Shawn S. , Liu Chenxi , O'Donoghue Anthony J. , LeibundGut-Landmann Salomé , Oon Hazel H. , Li Hao , Dawson Thomas L. 
Published: September 2022

Abstract

S1.3 Malassezia: genetics, genomics, and biology, September 21, 2022, 11:00 AM - 12:30 PM

Objectives
Malassezia forms the dominant eukaryotic microbial community on the human skin. The Malassezia genus possesses a repertoire of secretory hydrolytic enzymes involved in protein and lipid metabolism which alter the external cutaneous environment. The exact role of most Malassezia secreted enzymes, including those in interaction with the epithelial surface, is not well characterized.

Methods and Results
In this study, we compared the expression level of secreted proteases, lipases, phospholipases, and sphingomyelinases of M. globosa in healthy subjects and seborrheic dermatitis or atopic dermatitis patients. We observed upregulated gene expression of the previously characterized secretory aspartyl protease MgSAP1 in both the lesional and non-lesional skin sites of affected compared to healthy subjects. To explore the functional roles of MgSAP1 in skin disease, we generated a knockout mutant of the homologous protease MfSAP1 in the genetically tractable M. furfur. We observed the loss of MfSAP1 resulted in dramatic changes in the cell adhesion and dispersal in both culture and a human 3D reconstituted epidermis model. In a murine model of Malassezia colonization, we further demonstrated MfSAP1 contributes to inflammation as observed by reduced edema and myeloid pustule formation with the knockout mutant versus wildtype.

Conclusion
Taken together, we show that this dominant secretory M. aspartyl protease has an important role in enabling a planktonic cellular state that can potentially aid in colonization and additionally as a virulence factor in barrier-compromised skin, further highlighting the importance of considering the contextual relevance when evaluating the functions of secreted microbial enzymes.
S10.4c Successful treatment of mucormycosis in hematological diseases
Authors: Kakeya Hiroshi 
Published: September 2022

Abstract
The diagnosis of mucormycosis relies upon the identification of organisms in tissue by histopathology with culture confirmation. However, culture often yields no growth, and histopathologic identification of an organism with a structure typical of Mucorales may provide the only evidence of infection. PCR-based technique may contribute to the early diagnosis of mucormycosis. We developed a new antigen test. We searched for secreted or membrane-bound proteins of Rhizopus oryzae. Protein RSA (Rhizopus-specific antigen, 23 kDa) was detected at significantly higher concentrations in serum and in lung homogenates of the R. oryzae-infected mice as compared to those of uninfected mice. And we will show a case of hematological disease with diagnosis using RSA test and successful treatment with Liposomal amphotericin B. Our study indicates that protein RSA may be a promising biomarker of R. oryzae infection.
S2.2a African histoplasmosis
Authors: Oladele Rita 
Published: September 2022

Abstract

S2.2 Histoplasmosis and talaromycosis, September 21, 2022, 3:00 PM - 4:30 PM

African histoplasmosis caused by Histoplasma carpsulatum var dubosii is endemic in Africa with few cases reported from outside Africa usually attributed to travelling or visits to sub-Saharan Africa. The pathogenesis is yet unclear. Infection can be acquired via inhalation of microconidia or by direct inoculation. African histoplasmosis commonly presents with papules, nodules, ulcers, swellings, lymph node enlargement, eczematoid, or psoriosiform skin lesions. Subcutaneous abscesses may also develop with discharging sinuses containing yeast cells of the fungus. Although it is generally believed to be acquired through inhalation, the lungs are usually spared. Disseminated forms are usually characterized by the involvement of bones and other organs including the gastrointestinal tract. As a result of limited availability of diagnostics, data on its prevalence and epidemiology are scarce. As with classical histoplasmosis, African histoplasmosis also mimics other clinical entities including TB and neoplasms. More awareness and a high index of suspicion on the part of clinicians will lead to early diagnosis and invariably improve clinical outcomes. An extensive review of literature revealed 365 cases of African histoplasmosis reported globally; 236 cases from Africa and 38 cases from other geographical regions including an indigenous case report from India; the location of the remaining cases was not found, positive HIV status was found in 75 cases only. No statistically significant relationship was observed when comparing the relationship between positive HIV status and fatal outcomes (P &gt;.05, Fisher's exact test). A case report from the UK mentioned 21 cases of African histoplasmosis previously identified in the region and all were associated with travelling to Africa. Out of the 365 identified cases, diagnostic modality was specified in 264 cases; histopathology (87.9%, n = 232), culture (20.8%, n = 55), microscopy (7.2%, n = 19), serology (2.7%, n = 7), cytology (n = 1, 0.4%), polymerase chain reaction (n = 43, 16.3%), and peripheral blood film (n = 1, 0.4%). Amphotericin B (n = 53), itraconazole (n = 37), and ketoconazole (n = 26) were the predominant antifungals used for treatment. More studies are required to ascertain the true burden and epidemiology of African histoplasmosis and to determine whether these cases reported outside Africa were autochthonous or imported from Africa. Diagnosis by culture, although the gold standard is also not routinely achieved due to lack of biosafety level 3 cabinet especially in resource-limited settings, leading to significant under diagnosis. It is imperative that capacity building and strengthening is instituted to aid the diagnosis and management of African histoplasmosis. There is a need for more studies to ascertain whether there are additional phylogenetic species within the African clade.
S3.2 Challenging clinical cases
Authors: Singhal Tanu , Gopalakrishnan Ram , Soman Rajeev N. , Patel Atul 
Published: September 2022

Abstract

S3.2 Challenging clinical cases, September 21, 2022, 4:45 PM - 6:15 PM
A case of hepato-splenic-renal candidiasis will be discussed with the aim to track down the causative organism and its likely susceptibility. The role of BDG, selection of the antifungal agent, and duration of therapy will be discussed.

A case of an HIV-infected patient with cryptococcal meningitis, pulmonary infection, and pulmonary IRIS will be discussed.

A case of Candida auris fungemia and organ infection in a liver transplant recipient who developed serially increasing MIC will be presented. The relevance of MIC, tentative breakpoints, and selection of antifungal agents based on PK PD considerations will be discussed.

Talamycosis is usually seen in HIV-infected individuals from North East India. Here we discuss a 3-year-old girl with acute lymphoblastic leukemia from Western India who developed cavitary pneumonia and multiple brain abscesses due to talaromyces. The case posed significant management challenges pertaining to both the fungal infection and cancer.

Renal mucormycosis in healthy immunocompetent hosts is an entity generally reported only from India and China. We discuss the case of a previously healthy 3-month-old infant who presented with acute renal failure and renal mucormycosis. The case is unique with respect to the source of infection and management strategy.

Treatment of fungal infections involving the draining system of the kidney is challenging mainly due to limited antifungal drug penetration at the site of infection. Challenges in the treatment of amphotericin-resistant Aspergillus terreus pelvicalyceal infections will be discussed.

CAPA is a well-known complication of coronavirus disease 2019 (COVID-19), but usually involves the lung alone and rarely disseminates when treated. This case illustrates a rare complication of COVID-19, namely endocarditis caused by Aspergillus.

Rhinosporidiosis is a well-known disease that usually involves mucosal surfaces in the head and neck. This case illustrates a rare occurrence, namely dissemination to multiple areas and mimicking the clinical presentation of other cutaneous mycoses.