bluuebunny/crossref_metadata_2025_split huggingface dataset:
P433 Identification of Candida species isolated from hospitalized patients with candiduria
Authors: Nabili Mojtaba , Moazeni Maryam 
Published: September 2022

Abstract

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

Background and objectives
The incidence of candiduria caused by Candida spp. has increased in recent years, particularly in hospitalized patients. Candiduria is most commonly caused by C. albicans; however, an increase in the prevalence of non-albicans species has been observed during last decades. This study aimed at molecular identification of Candida species isolated from candiduria in hospitalized patients.

Methods
This cross-sectional study was conducted on 530 hospitalized patients in two hospitals in the Mazandaran Province, Iran. Midstream urine specimens were collected and then cultured on CHROMagar Candida medium. Molecular identification of common Candida species was carried out using the polymerase chain reaction-restriction fragment length polymorphism method after enzymatic digestion with MspI. C. albicans and Candida parapsilosis species complexes were identified by amplification of the HWP1 and intein-containing vacuolar ATPase precursor genes, respectively.

Results
The frequency of candiduria was estimated at 14% among hospitalized patients. Of 74 samples positive for candiduria, 65 (87.8%) were isolated from females. The most common predisposing factor to candiduria was diabetes (n = 36; 48.6%). The most frequent isolates were C. albicans complex (n = 44; 59.4%), followed by C. glabrata (n = 16; 21.6%), C. tropicalis (n = 10; 13.5%), C. krusei (n = 3; 4%), and C. parapsilosis (n = 1; 1.3%).

Conclusion
Based on the results, the conventional and molecular methods produced similar results for the identification of Candida species. However, accurate identification of Candida spp. requires the use of molecular techniques such as PCR-RFLP, HWP1, and intein-containing vacuolar ATPase precursor genes. Nevertheless, chromogenic methods such as CHROMagar Candida can be used for the diagnosis of Candida spp. in laboratories with limited resources.
P447 Galactomannan lateral flow assay for the diagnosis of invasive Aspergillosis among clinically suspected patients in tertiary care center, Jodhpur, Rajasthan
Authors: Priya Hari , Kothari Nikhil , Jain Vidhi , Kumar Deepak , Khichar Satyendra , Rohila Amit Kumar , Mokta Mohan Lal 
Published: September 2022

Abstract

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

Introduction
Invasive aspergillosis is one of the potentially life-threatening diseases in immunocompromised patients. Early diagnosis and prompt treatment improve patient survival. The gold standard method—conventional microscopy and culture have low sensitivity and a long turnaround time. Serum Galactomannan (GM), a polysaccharide that forms a major component of Aspergillus cell wall and is released by the fungus during invasive growth is established as a reliable biomarker, which is available as Enzyme Linked Immunoassay (ELISA). The limitations of ELISA are high cost, expertise, and difficulty in assay standardization. To overcome these limitations, a qualitative Galactomannan Lateral Flow Assay (GM-LFA) a sandwich immunochromatographic test, recently approved by European CE, is evaluated in our study.

Objective
To establish the diagnosis of Invasive Aspergillosis according to EORTC/MSGERC Definitions of Invasive Fungal Diseases 2021 guidelines and comparison of GM-LFA with conventional microscopy and culture.

Method
We performed a retrospective study from October 2021 to March 2022 on serum and bronchoalveolar lavage fluid (BAL) samples. GM-LFA (IMMY sōna Aspergillus Galactomannan LFA) was performed and the results are read after 10 mins with a cube reader according to the manufacturer’s instructions and compared with conventional microscopy and culture.

Result
During the 6 months study period, 185 samples (14 BAL and 171 Serum) were collected from 148 patients. A total of 17/185 (9.18%) samples from 16 patients were positive by GM-LFA, of those 5 (29.41%) are BAL, and 12 (70.58%) are serum samples. One patient tested positive for Galactomannan on both BAL and serum.
Among Assay positive samples, 3/17 (17.64%) samples were positive by microscopy and culture, which grew 2 Aspergillus fumigatus and 1 Aspergillus niger.
Treatment details of 5 patients could be traced, of which 4 patients improved clinically and radiologically after Inj. voriconazole and 1 patient died before starting treatment.
According to EORTC/MSGERC Definitions of Invasive Fungal Diseases 2021 guidelines, 17 patients belong to the proven and probable Invasive aspergillosis category, in which 16 patients were GM-LFA-positive and the remaining 131 patients had no evidence of invasive aspergillosis disease.

Conclusion
GM-LFA is a cost-effective, easy, convenient, and rapid procedure. We recommend its use for the diagnosis of invasive Aspergillosis in routine healthcare settings, especially where GM-ELISA is not available or affordable.
S1.4b Challenges in diagnosing and management of invasive fungal infections during the pandemic
Authors: Chayakulkeeree Methee 
Published: September 2022

Abstract

S1.4 Fungal infections in Asia, bringing it out of the dark, September 21, 2022, 11:00 AM - 12:30 PM
Invasive fungal diseases have been increasing in Asian countries. Recent advances in novel medical care such as solid organ and stem cell transplantations, chemotherapy for cancer treatment, and corticosteroid therapy, resulted in the increased prevalence of invasive mycoses. Invasive aspergillosis, mucormycosis, and endemic mycoses are among the most common mold infections in Asia. Non-classical and novel risk factors of invasive fungal diseases have been increasingly recognized in Asia. In contrast to the classical neutropenic patients, most of the patients with invasive mycoses who had non-classical risk factors are mostly non-neutropenic and may present with an atypical clinical manifestation. These novel risk factors include biologic agents or small molecule kinase inhibitors used for cancer treatment, and severe viral pneumonia such as influenza pneumonia or coronavirus disease 2019 (COVID-19) pneumonia. Recently, COVID-19-associated aspergillosis (CAPA) and COVID-19-associated mucormycosis (CAM) have been described. These particular mold infections had high mortality. Treatment of CAPA and CAM are similar to those who had mold infections without COVID-19. However, the interaction between triazole and drugs used for the treatment of COVID-19 must be taken into consideration.

Timely and accurate diagnoses are crucial for the management of invasive fungal infections. Conventional fungal cultures from sterile clinical samples or blood are useful but they are time-consuming. Nevertheless, the diagnosis of invasive mold infections is challenging as the imaging is non-specific and the serological tests are not widely available in Asian countries. In some circumstances such as those with non-classical risk factors, serology revealed relatively low sensitivities. Molecular diagnostic tests are also the unmet needs among Asian countries for timely and accurate diagnosis of invasive fungal diseases. Several factors should be considered for the appropriate choice of antifungal agents, including antifungal coverage, adverse effects, underlying conditions, drug-drug interactions, and cost. Recently, novel antifungal agents such as novel triazoles or new classes of antifungal agents have been studied and may be a promising choice for the treatment of invasive fungal diseases.
S10.4a New mechanism and detection methods for azole-resistant Aspergillus fumigatus
Authors: Watanabe Akira , Arai Teppei , Majima Hidetaka 
Published: September 2022

Abstract

S10.4 Emerging antifungal resistant fungi, September 24, 2022, 10:30 AM - 12:00 PM
The most studied azole-resistant mechanism of Aspergillus fumigatus is decreased affinity of the drug for CYP51A, the drug target molecule, due to its amino acid substitutions. Typically, each azole-resistance caused by the designated amino acid substitution of CYP51A has a specific pattern depending on the substitution site. While uncovering non-cyp51A mechanisms responsible for azole resistance should be essential for developing novel methods for prompt diagnosis and effective drug treatment. In our previous study, we reported results that mutation of hmg1, which codes HMG-CoA reductase, the rate-limiting enzyme in ergosterol biosynthesis, would be the mechanism conferring azole-drugs resistance (EID 2018). On the other hand, different azole susceptibility patterns have been reported even among the strains possessing the same mutation in CYP51A. In this way, the overall picture of molecular mechanisms inducing azole resistance remains unclear.

We performed a comparative genomic analysis among the strains with the same cyp51A mutation isolated from the same patient but with different azole resistance patterns. To investigate the association between the novel mutation and azole resistance, the mutant allele was replaced with the wild-type allele by the CRISPR-Cas9 system. Antifungal susceptibility tests were performed according to the CLSI-M38.

As a result of genome comparison analysis between these two strains using the HiSeq sequencing system (Illumina), another mutation was found in the insulin-induced protein (INSIG) of the multiazole-resistant strain. The INSIG mutation contributes additively to azole resistance in collaboration with the CYP51A mutation but does not alone itself.

We have already reported simple and rapid detection methods for A. fumigatus possessing CYP51A mutation using an endonuclease (AAC 2020). Furthermore, using MALDI-TOF-MS, we are developing a discriminant model to detect azole-resistant A. fumigatus.
S2.3d Mixed biofilm of Aspergillus fumigatus and Stenotrophomonas maltophilia: Microscopic visualization of galactosaminogalactan and galactomannan polysaccharides in the extracellular matrix
Authors: Valsecchi Isabel , Debourgogne Anne , Fontaine Thierry , Botterel Françoise 
Published: September 2022

Abstract

S2.3 Novel diagnostic tools for invasive mold infection, September 21, 2022, 3:00 PM - 4:30 PM

Introduction
Aspergillus fumigatus (Af) and Stenotrophomonas maltophilia (Sm) are commonly co-isolated from the airways of cystic fibrosis patients, especially in mixed biofilms. A mixed Af-Sm biofilm model, developed by our lab, demonstrated that Sm exhibits an antibiosis effect on Af by (1) inhibiting fungal growth, (2) rendering the fungal mycelia highly branched, and (3) increasing the fungal cell wall thickness.1 The presence of three Af cell wall polysaccharides, galactomannan (GM), galactosaminogalactan (GAG), and α-1,3-glucan, have been described in the extracellular matrix (ECM) of Af biofilms.2 GM and GAG are known to be released out from the cell wall of Af.3 GAG contains N-acetylgalactosamines (GalNAc) which undergo partial deacetylation extracellularly, to be converted to GalN, and thus transform this polysaccharide into a polycation with strong surface binding properties.4

Aim of the study
Analyze the structure of fungal cell wall polysaccharides in the formation of Af biofilm alone or mixed with Sm.

Methods
The Af-Sm co-culture was prepared on an 8-chambers Lab-Tek slide as described previously1. Anti-GAG and anti-GM monoclonal antibodies produced at the Aspergillus Unit at Pasteur Institute of Paris, were used to focus on the 3D structure of GAG and GM in the ECM of the biofilm. These polysaccharides were analyzed by fluorescence confocal microscopy at 24 h.

Results and Conclusion
GM was found, as expected, in the hyphal Af cell wall but was very little in the ECM. GAG was also found in the cell wall but mainly formed a beautiful fibrillary network between the hyphae, showing the importance of this polysaccharide in cell-cell interaction and in the structuration of Af biofilm. GAG could be the surface receptor for Sm, which would promote strong adhesion between Sm and Af in the biofilm.

Sources
1. Melloul E et al. Front Microbiol. 2018;
2. Beauvais A et al. Microbiol Spectr. 2015;
3. Fontaine T et al. J Fungi. 2020;
4. Briard B et al. Mycologia 2016.
S3.1a Update in mycetoma control
Authors: Fahal Ahmed Hassan 
Published: September 2022

Abstract

S3.1 Neglected implantation mycoses, September 21, 2022, 4:45 PM - 6:15 PM
Mycetoma is a badly neglected tropical disease, reported globally but endemic in many tropical and subtropical, and Sudan has the highest disease burden. Mycetoma epidemiological features remain uncharacterized. Hence, there is no preventive or control program for mycetoma worldwide. To reduce the disease burden and its socio-economic impacts on patients, families, communities, and the health system in the endemic regions, the Mycetoma Research Center, University of Khartoum, Sudan, WHO Collaborating Center on Mycetoma has adopted a unique holistic approach to management and mycetoma patients, and it gained a great experience on that.

In this presentation, we will share the MRC's experience in dealing with the patients at the endemic villages level, supporting the affected communities and collaborating with the local health authorities to improve the life quality of the affected patients.
S3.3c Alveolar macrophage-mediated host resistance against Aspergillus fumigatus
Authors: Obar Joshua 
Published: September 2022

Abstract

S3.3 Innate immune responses to pathogenic fungi, September 21, 2022, 4:45 PM - 6:15 PM

Alveolar macrophages (AlvMφ) reside on the luminal surface of the airways serving as the primary phagocyte within the airways of the lungs where they act as immune sentinel cells sensing and responding to microbial and environmental exposures. In this role, AlvMφ must be able to respond in a manner that is appropriate to the threat posed which has been hypothesized to occur through sensing microbial vitality and/or patterns of pathogenesis. It is well-established that AlvMφ interact with phagocytose and respond to A. fumigatus, but their role in host resistance against A. fumigatus is currently controversial. Here I will discuss the role of AlvMφ play in orchestrating a robust and effective antifungal innate immune response to mediate A. fumigatus clearance. AlvMφ orchestrate the protective innate immune response against A. fumigatus by sensing live fungal conidia using the cytosolic RNA-sensing MDA5 receptor to initiate the host protective type I and type III interferon response in both mice and humans. The activation of MDA5/MAVS signaling appears to be mediated by both fungal dsRNA-dependent and fungal dsRNA-independent mechanisms. Thus, AlvMφ serve as a central hub for regulating and tuning the antifungal immune response within the respiratory tract.
S3.4b Lactoferrin, a natural source of peptides that potentiate the antifungal activity of amphotericin B
Authors: Fernandes Kenya , Desplazes Evelyne , Kulkarni Sameer , Payne Richard , Carter Dee 
Published: September 2022

Abstract

S3.4 Free oral paper session, September 21, 2022, 4:45 PM - 6:15 PM

Objectives
It is notoriously difficult to prevent and treat fungal infections, however, the natural world has come up with remedies that are non-toxic, effective, and evade resistance. Here we investigate lactoferrin, an iron-binding glycoprotein found in milk, tears, and sweat, for its capacity to inhibit fungi and to synergize with commonly used antifungal drugs, with the aim of determining its mode of action.

Methods
Lactoferrin (LF) was obtained from a commercial supplier and two dairy companies. LF was tested on suite of pathogenic yeast and mold species for inhibition using CLSI microdilution methods. Synergy was determined with antifungal drugs amphotericin B (AMB), nystatin (NYS), fluconazole (FLC), itraconazole (ITC), voriconazole (VRC), and 5-fluorocytosine (5FC). The effect of LF on fungal cells was analyzed using scanning electron microscopy (SEM). The active peptide/s within LF were then predicted from pepsin and in silico digestion, synthesized, and tested for synergy with amphotericin B (AMB). Tethered synthetic membranes were produced and were loaded with ergosterol or cholesterol to test the nature and specificity of membrane binding by LF and the synthetic peptide.

Results
LF demonstrated antifungal activity against yeast species Cryptococcus, Candida, and Saccharomyces and was much less effective against molds. Good synergy was achieved with AMB but not azole or echinocandin drugs. While the iron-chelating capacity of LF was important for the antifungal activity it was not involved in synergy. SEM revealed cell damage suggesting an interaction between AMB, LF and the fungal membrane or cell wall. A 30-residue peptide from the C lobe of LF was synthesized and tested for activity and synergy. This peptide, dubbed lactofungin (LFG) was inactive alone but was potently synergistic with AMB, indicating a direct role in augmenting AMB activity. Synthetic membranes loaded with ergosterol but not cholesterol were disrupted by AMB + LFG, demonstrating that activity was fungal-specific and was mediated through ergosterol binding.

Conclusion
LF is a complex molecule that causes fungal inhibition via iron binding and when cleaved by pepsin can produce active peptides. As AMB is a highly toxic treatment, the use of LFG as a synergent could help increase activity while lowering the effective dose, thereby reducing undesirable side effects. The action of AMB + LFG appears dependent on ergosterol, suggesting inhibition will be highly fungal-specific.
S3.4c A pipeline toward the identification of novel antifungal compounds derived from the microbial dark matter
Authors: Vanreppelen Giel , Wuyts Jurgen , Holderbusch Alannah , Peys Alexander , Hu Haibo , De Ruysscher Dries , Masschelein Joleen , Luyten Walter , Vandecruys Paul , Van Dijck Patrick 
Published: September 2022

Abstract

S3.4 Free oral paper session, September 21, 2022, 4:45 PM - 6:15 PM

Background
The current armamentarium of antifungal drugs and the restricted variety in antifungal drug classes combined with the ever-rising threat of resistant fungal pathogens highlighted the urgent need for novel antifungal compounds. Natural antifungal secondary metabolites have always been the prevalent source for drug development, exemplified by the echinocandin and polyene drug classes. Yet, the golden age discovery platforms were abandoned due to compound rediscovery and its paired economic cost.

Study
In an effort to revive the original success stories, we combined the traditional approach of sampling and screening for active secondary metabolites with present-day advances in sequencing, genome mining, impedance spectroscopy, HPLC, LCMS, and NMR.
Soil bacteria and fungi were isolated through in situ cultivation via the iChip method. After application of the OSMAC approach, 389 broths were identified with activity against Candida albicans. To prioritize active extracts, several criteria were set up; low to absent mammalian host cell toxicity, activity against a broad spectrum of fungal pathogens including wild-type reference strains, and established antifungal drug-resistant variants and species identification of the producing strain. Continuing, Lead hits were purified utilizing bioactivity-based semi-preparative HPLC. The resulting pure fractions were analyzed by tandem LCMS-MS, and proposed structures were later confirmed with NMR. In vitro and in vivo validation of the purified compounds will be performed.
Additionally, aside from discovering a novel antifungal compound, another project goal is to gauge if impedance spectroscopy can provide an early suggestion regarding the mode of action of the present antifungal agent. For this, a POC study was performed which showed that different antifungal drug classes provide distinct signature response profiles by which they can be classified. As such, when active secretion broths show unique impedance profiles, in comparison with the signature profiles of established antifungal drugs, it suggests that they work through a different mode of action.

Results
Several species were identified as producing antifungal secondary metabolites that are currently absent in the literature. Either the compound itself was unknown or literature never described the species as a producer of a known, or variant of a known antimycotic compound. Moreover, several species are novel based on Illumina sequencing. Genera producing our current lead hits include bacteria: Pseudomonas, Tsukamurella, Paraburkholderia, and fungi: Athelia, Penicillium. Within the collection, the Pseudomonas species appear to produce variants of the antimycotic iron-chelating pyoverdine class.
S4.1a Fusariosis: MICs, mono versus combination therapy and fosmanogepix
Authors: Hoenigl Martin 
Published: September 2022

Abstract

S4.1 Treatment of rare mold infections in 2021: the role of new and old antifungals, September 22, 2022, 10:30 AM - 12:00 PM
Fusarium is one of the most clinically prevalent rare molds causing superficial infections such as keratitis in immunocompetent hosts and severe disseminated infections frequently presenting as fungemia in the immunocompromised. These fungi are ubiquitous in nature and are found in soil and air. Only a few of the ˃ 70 Fusarium spp. are opportunistic pathogens in humans. F. solani and F. oxysporum species complexes are specifically important, causing ˃50% and about 20% of severe fusariosis cases, respectively. Although galactomannan and also beta D glucan may result positive in fusariosis, fungal culture from either blood, BAL, or skin biopsy remains the most frequently utilized test for diagnosing fusariosis. Susceptibility testing of isolates recovered in culture should be used for epidemiologic purposes in defining the range of minimal inhibitory concentrations (MIC) distribution of Fusarium spp., however, studies that prove that susceptibility testing results should be utilized to inform antifungal drug choice are lacking. Although there are no interpretive breakpoints for antifungal agents against Fusarium spp., compounds with MICs that are off-scale, such as &gt;16 μg/ml, at the highest range of concentrations are unlikely to be active in vivo or in patients. Initial combination therapy with a lipid formulation of amphotericin B plus voriconazole, deescalated to monotherapy once MICs come back, is, therefore, a frequently chosen management pathway.

A number of drugs in the antifungal pipeline will present additional treatment options for Fusarium, including Fosmanogepix and Olorofim.