bluuebunny/crossref_metadata_2025_split huggingface dataset:
P341 Evaluation of anti-Candida activity of Cinnamaldehyde using mice model
Authors: Uttam Gunjan , Singh Karuna 
Published: September 2022

Abstract

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

Objective
Vaginal candidiasis is a frequent infection afflicting the female population. Candida co-infections are reported in escalating frequency and can be associated with severe health hazards or even death. People with severe COVID-19, malignancies, AIDS, and organ transplants are particularly vulnerable to invasive vaginal candidiasis. Due to the high frequency of infections associated with recurrence, vaginal candidiasis poses a significant medical problem worldwide. Treatments of vaginal candidiasis are limited due to drug resistance, side effects, and toxicity. CIN is a natural compound and its antifungal activity is widely reported. The introduction of cinnamaldehyde (CIN) as the anti-Candidal agent will revolutionize the treatment of vaginal candidiasis.
In this study, we investigated the anti-Candida activity of CIN against vaginal candidiasis in Swiss albino mice (C3HHC-Strain).

Methods
Vaginal candidiasis in mice (Swiss albino) was induced under conditions of pseudo-estrus. Persistent vaginal infection was found in estrogenized mice after vaginal challenge with C. albicans. The mice were treated orally after confirmation of infection in mice. The efficacy of CIN treatment was investigated phenotypically by colony-forming unit (CFU) counts in the vaginal smear, fungal load determination in the blood, the ovarian and vaginal tissues and periodic acid-Schiff (PAS) staining of histopathological sections of the vaginal tissues. The hematological parameters of the experimental mice were also evaluated.

Results
The pseudohyphae and spores of C. albicans were present in the vaginal smear of experimentally infected mice. After treatment, no C. albicans colonies were found in the vaginal lavage of infected mice. The fungal burden was significantly higher in the vagina and the ovaries of infected mice. However, a dose of 262.5 mg/kg BW of CIN reduced the number of CFU in the vagina and ovaries in treated mice. The histopathology revealed the absence of C. albicans in the vaginal tissue of the treated mice. Nonetheless, the vaginal sections of infected mice exhibited pathological changes. The hematological parameters such as RBC count, WBC count, and percentage of hemoglobin showed significant differences in the treatment groups compared to the infected group.

Conclusion
Cinnamaldehyde showed good in vivo antifungal potential against vaginal candidiasis. However, evaluation of its pharmacodynamic and pharmacokinetic parameters and complete elucidation of its mode of action are desirous.
P349 Relationship between genotypes and in vitro pathogenic behavior of Uruguayan strains of Cryptococcus spp.
Authors: Cabeza Elisa , Arteta Zaida , Betancor Laura 
Published: September 2022

Abstract

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

Objective
Contribute to the knowledge of the relationship between genetic and pathogenic characteristics of meningeal cryptococcosis-producing strains from Uruguay.

Methods
We have included a total of 21 isolates of Cryptococcus spp from human cases of meningoencephalitis. We start from the general strain collection of the mycology laboratory of the department of parasitology and mycology, medical school, University of the Republic. The strain was sectioned taking into account the phenotypic diversity and genetic diversity through a previous analysis by PCR fingerprinting, for study by MLST.
MLST was performed according to the protocol available at: http://mlst.mycologylab.org/defaultinfo.aspx?Page=CryptoHome.
Infection, intracellular survival, and cell proliferation assays were performed. The murine macrophage cell line J774 (ATCC TIB 67) was used.

Results
Of the 21 strains studied, in 13 we obtained an ST that had been previously reported (9 C. neoformans complex and 4 C. gattii complex). For the remaining 8 strains, we obtained new allelic combinations to which new STs were assigned, which were deposited in the MLST database for Cryptococcus spp. (http://mlst.mycologylab.org) (Table 1).
All strains were phagocytosed and all were able to persist within macrophages at 24 h post-infection, although there was great variability between strains (Table 1). In most of the strains, the control of macrophages prevailed through cell lysis mechanisms over Cryptococcus spp, with one proliferation index (PI) ˂1.0. In the proliferation assay, only 4 strains managed to proliferate within the macrophages, 3 of them correspond to C. gattii complex, genotypes VGI and VGII. The 4th strain that achieved proliferation was corresponding to the VNI genotype.

Conclusion
We obtained a clear differentiation of in vitro behavior between the strains corresponding to C. neoformans complex and C. gattii complex. The strains belonging to C. gattii complex presented less phagocytosis and greater persistence and intracellular proliferation.
P384 Characterization of the virulence potential of Aspergillus species of section Terrei in Galleria mellonella infection model
Authors: Shahandashti Roya Vahedi , Binder Ulrike , Houbraken Jos , Lass-Flörl Cornelia 
Published: September 2022

Abstract

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

Objectives
Species belonging to the genus of Aspergillus are among the most common causative agents of human and animal infections. Less than 40 species among all Aspergillus species are known to be associated with human infections, including allergic bronchopulmonary aspergillosis, chronic pulmonary aspergillosis, and invasive aspergillosis. And of these, Aspergillus section Fumigati is one of the major infectious causes of death, followed by members of sections Flavi, Nigri, and Terrei. Aspergillus species in section Terrei are categorized into three series: Ambigui, Nivei, and Terrei. A. terreus sensu stricto is the first species described and the most common species found worldwide in different ecological habitats.
However, there are several other species within the section Terrei, stating cryptic species which are not distinguished by conventional morphological analysis, even though they are taxonomically accepted by forming a distinctive phylogenetic clade. Despite definitive species identification, there is still less known about the virulence potential of all species in this section, and it might be underestimated because of their lack of distinction by conventional diagnostic methods. In this ongoing study, the in vivo Galleria mellonella model has been utilized to examine the inter-and/or intraspecies virulence dependency of section Terrei.

Methods
A total of 18 accepted Aspergillus species in section Terrei (n = 18) were tested, including A. terreus sensu stricto, A. citrinoterreus, A. hortae, A. pseudoterreus, A. alabamensis, A. aureoterreus, A. floccosus, A. iranicus, A. recifensis, A. carneus, A. microcysticus, A. niveus, A. bicephalus, A. neoindicus, A. neoafricanus, A. barbosae, A. ambiguus, and A. allahabadii. Species were identified by sequencing gene regions of b-tubulin, calmodulin, and RNA Polymerase II Subunit 2 (RPB2). Briefly, groups of larvae (n = 30) (0.3 to 0.4 g; SAGIP, Italy) were stored in wood shavings in the dark at 18°C for 24 h before the experiment. Three groups were included: larvae infected with 107 conidia/larva, larvae injected with 20 μL sterile insect physiological saline, and untouched larvae. The survival rate was monitored for up to 144 h at 37°C.

Results
Median survival rates revealed a species-dependent virulence pattern. Larvae inoculated with A. aureoterreus, A. pseudoterreus (Serie Terrei) and A. niveus, A. carneus, and A. iranicus (Serie Nivei) exhibited high virulence potential by reflecting lower survival rates in comparison with other species. In contrast, species belonging to the series Ambigui showed low virulence potential.

Conclusion
In conclusion, the virulence characteristics of section Terrei differ between species. Further studies are needed to unravel the species' invasiveness, such as histopathology and immune response of G. mellonella.
P396 Utility of nasal swabs as samples for the diagnosis of Mucormycosis using molecular methods for detection
Authors: Chowdhury Souradeep , Sikdar Sunit , Ranjan Piyush , Singh Gagandeep , Pandey Mragnayani , Ghosh Tamoghna , Sachdev Janya , Yadav Renu , Perumalla Sowjanya , Bhattacharya Aakashneel , Xess Immaculata , Wig Naveet 
Published: September 2022

Abstract

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

Objectives
Mucormycosis is an aggressive, life-threatening infection caused by fungi in the order Mucorales. There was an explosion of new cases of rhino-sino-orbital mucormycosis following the COVID pandemic in India, and the need for easy and rapid diagnostics was felt. The current diagnosis of mucormycosis relies on mycological cultures, radiology, and histopathology. These methods lack sensitivity and are most definitive later in the course of infection, resulting in the failure of timely intervention.
A real-time multiplex PCR platform is commercially available for the detection of Rhizopus spp., Mucor spp. Rhizomucor spp., Lichtheimia spp., and Cunninghamella spp. (PN-700, MucorGenius®, PathoNostics®, Maastricht, The Netherlands) This real-time PCR has been validated to identify these fungal pathogens from bronchoalveolar lavage, tissue, and serum samples. This study aimed to validate this PCR-based system to detect Mucorales from nasal swab samples and evaluate its utility in the detection of Mucorales from nasal cavities of high-risk patients developing signs and symptoms of mucormycosis.

Methods
A single-center cross-sectional observational study was conducted on 50 hospitalized adult patients with signs and symptoms of mucormycosis. Nasal swabs were taken for PCR analysis once there was a clinical suspicion and were compared with the results of the gold standard. The gold standard for the diagnosis of mucormycosis was the conventional method (KOH mounted microscopy/HPE). Demographic details and risk factors for these patients were recorded, and the RTPCR-based test was run on the nasal swab samples of all these 50 patients. The workflow is depicted graphically in Fig. 1 (Created with BioRender.com).

Results
The study population mean (SD) age was 50 (16) years and consisted of 32 (64%) males. A total of 39 (78%) patients were known cases of diabetes mellitus, 48 (96%) patients had amphotericin B intake, and 20 (40%) had posaconazole intake. In all, 21 (42%) patients had a past history of COVID-19 infection; 14 patients had received steroids and 10 patients received oxygen support. PCR for Mucorales was positive in 15 (30%) patients while the KOH mount was positive in 4 (8%) patients.

Conclusion
These results are not encouraging for the use of nasal swabs as the sample for diagnosis of mucormycosis. Though the PCR performed better on the swab samples than KOH preparation and culture techniques, it highlights the importance of using standard sampling methods.
P4 COVID19-associated fungal infections
Authors: Hoenigl Martin 
Published: September 2022

Abstract

Plenary session 4, September 22, 2022, 2:00 PM - 2:45 PM

Coronavirus disease 2019 (COVID-2019) associated invasive fungal infections (IFIs) have emerged as an important complication in a substantial number of critically ill COVID-19 patients, and three groups of fungal pathogens have increasingly been recognized as causes of superinfections: Aspergillus, Mucorales, and Candida. First reports of cases and case series of COVID-19-associated pulmonary aspergillosis (CAPA) have emerged during the first months of the pandemic. Prevalence rates varied widely due to the fact that CAPA was, and still remains, challenging to diagnose in patients with COVID-19-associated acute respiratory failure (ARF). The clinical picture and radiological findings of CAPA are unspecific and the primarily airway invasive growth in non-neutropenic patients and the late occurrence of angioinvasion in the course of the disease may complicate diagnosis. Current guidelines recommend treatment of CAPA during its early airway invasive phase, which may result in some overtreatment (i.e., treatment in patients that may not develop angioinvasive infection), given the independent contribution of CAPA to devastating mortality rates of around 50% that have been shown in multiple studies. This talk will also review the incidence of COVID-19-associated IFIs caused by Mucorales, and Candida, including C. auris, and will discuss—clinical risk factors, predisposing changes in the host environment, and immunological mechanisms involved in the pathogenesis of these coinfections, as well as current and future diagnostics and treatments.
P402 Comparison of PCR-RFLP with 21-plex PCR and rDNA: sequencing for identification of clinical yeast isolates
Authors: Khodavaisy Sadegh , Kord Mohammad , Elmimoghaddam Ahmad , Hashemi Seyed Jamal , Reziae Sassan , Ghazvini Roshanak Daie , Salehi Mohammadreza , Abdollahi Alireza , Ahmadi Ali , Boekhout Teun 
Published: September 2022

Abstract

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

Objectives
Non-albicans Candida species and other rare yeasts have emerged as major opportunistic pathogens in fungal infections. Identification of opportunistic yeasts in developing countries is mainly performed by phenotypic assay, which is time-consuming and prone to errors. The aim of the present study was to evaluate PCR-RFLP as a routinely used identification technique for the most clinically important Candida species in Iran and make a comparison with a novel multiplex PCR, called 21-plex PCR.

Methods
A total of 173 yeast isolates from clinical sources were selected and identified with sequence analysis of the D1/D2 domains of rDNA (LSU rDNA) sequencing as the gold standard method.

Results
The results were compared with those obtained by PCR-RFLP using MspI restriction enzyme and the 21-plex PCR. PCR-RFLP correctly identified 93.4% of common pathogenic Candida species (C. albicans, C. glabrata, C. parapsilosis, C. tropicalis, and P. kudriavsevii (=C. krusei)) and was able to identify 45.5% of isolates of the uncommon yeast species compared to the D1/D2 rDNA sequencing. Compared with PCR RFLP, all common Candida species and 72.7% of uncommon yeast species were correctly identified by the 21-plex PCR.

Conclusion
The application of the 21-plex PCR assay as a non-sequence-based molecular method for the identification of common and rare yeasts can reduce turnaround time and costs for the identification of clinically important yeasts and can be applied in resource-limited settings.
P403 Metagenomic sequencing as an effective diagnostic tool for Talaromycosis in HIV-negative patients: A retrospective study
Authors: Liang Tianwei , Cao Cunwei 
Published: September 2022

Abstract

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

Objective
The diagnosis of Talaromyces marneffei (T. marneffei) infection in HIV-negative patients is still challenging. The aim of this study was to evaluate the effectiveness and efficiency of metagenomic next-generation sequencing (mNGS) for T. marneffei infection in non-HIV-infected patients by comparing the diagnostic value of mNGS and traditional microbiological culture on different specimens.

Methods
A total of 57 HIV-negative patients with suspected infectious diseases were enrolled in the study from July 2018 to November 2021. mNGS and culture were performed on all enrolled samples. Data were collected on demographic characteristics, clinical symptoms, laboratory findings, and antifungal results. The diagnostic value of mNGS and conventional microbiological culture was compared.

Results
A total of 66 samples from 57 patients were analyzed, 29 were diagnosed with T. marneffei infection and 28 were of non-T. marneffei infected patients. Compared with the final diagnosis, mNGS showed a sensitivity of 97.22%, which was higher than that of conventional culture (61.11%). In addition, mNGS provides greater diversity in sample selection compared to culture.

Conclusion
Our study demonstrated that mNGS can be considered a promising tool for rapid and accurate pathogenic diagnosis in HIV-negative patients with suspected T. marneffei.
P423 Recombinase polymerase amplification for Mucormycosis
Authors: Hallur Vinaykumar , Sable Mukund , C. Preetam , Sahu Supriya , Tudu Malati 
Published: September 2022

Abstract

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

Objectives
(1) To develop a Recombinase polymerase amplification (RPA)-based assay to diagnose mucormycosis. (2) To determine the analytical sensitivity and cross-reactivity of the developed assay. (3) To validate the assay in clinical samples. (4) To determine the turnaround time for the developed assay.

Methods
Multiple alignments of the ITS region of Rhizopus arrhizus were used to identify conserved sequences and design 30-32 bp long primers and 45 bp long Exo probes for RPA-based assay as per TwistDX guidelines (Cambridge, UK) using the PrimedRPA program (Higgins et al. 2019) and subject to in silico specificity check using NCBI primer blast. Genomic DNA and plasmid clone of the target region were spiked into human DNA and were used to determine analytical sensitivity. DNA from 35 clinically relevant fungi and bacteria were used to determine the cross-reactivity. A total of 40 sinus tissue samples from patients with microscopy and culture-confirmed mucormycosis and 40 sinus tissue samples from patients without mucormycosis were used to determine the diagnostic sensitivity and specificity of the assay. DNA from relevant fungi, bacteria and molds were extracted using the MPbio FastDNA kit as per the manufacturer’s instructions with an elution volume of 100 μl. All extracted DNA samples were subjected to GAPDH PCR before RPA testing to rule out the presence of inhibitors as per established protocols. A 50 μl RPA reaction using Twistamp Exo kit as per manufacturer's instruction but using 10 μl template instead of 1 μl. The reactions were set up and inserted into a portable Qiagen ESE quant TS2.4 fluorometer incubated at 39°C and fluorescence was acquired every 30 sec over 20 min. The tubes were removed from the fluorometer mixed well and reinserted after 4 min as per kit instructions. Positive control and no template control were set up with each reaction. All relevant data were entered in Microsoft excel and diagnostic sensitivity and specificity were calculated using the Medcalc free online statistical software.

Results
A total of 1246 primer-probe combinations were designed and checked for in silico specificity using NCBI primer blast. Of these 1246 primer probes, 8 primers and 1 probe was finalized for in vitro testing to determine the best primer and probe set. The finalized primers had analytical sensitivity of 10pg and 10 copies and showed no cross-reactivity with any of the 35 clinically relevant fungi and bacteria. All extracted DNA showed amplified GAPDH gene demonstrating the absence of PCR inhibitors in extracted DNA. The sensitivity and specificity of the assay were 97.5% (95% CI: 86.8-99.9) and 97.6% (95% CI: 87.1-99.9). The turnaround time for RPA was 6 h.

Conclusion
Real-time RPA can be used to rapidly and reliably diagnose mucormycosis.
P443 Altered expression of fungal CotH, human glucose-regulated protein 78 (GRP78), and predicted miRNAs in macrophages and model diabetic mice infected with Rhizopus oryzae
Authors: Shokohi Tahereh , Seifi Zahra , Shafie Mohammad , Mowla Javad , Niknejad Farhad , Abastabar Mahdi , Larijan Lale Vahedi 
Published: September 2022

Abstract

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

Objectives
Rhizopus oryzae is one of the most common causes of mucormycosis. Among the virulence factors of the Mucorales, CotH protein has recently been identified, which causes the invasion of R. oryzae into endothelial cells. In this study, we aimed to examine the reaction between GRP78 at the level of human cells and different groups of mice and CotH3 at the surface of the R. oryzae hyphae. We evaluated the relative expression of GRP78 and CotH3 genes and changes in the expression of some miRNAs that target the human GRP78 gene.

Methods
In this study, the relative changes in gene expression were studied. In three groups (1) Macrophages derived from human monocytes: monocytes from the blood of healthy donors were isolated using Ficoll and in RPMI 1640 medium containing FCS 10% and with penicillin-streptomycin after 2 weeks were differentiated into macrophages. Two groups were investigated, including control and infected with R. oryzae hyphae, for 6 and 16 hours after infection. (2) Hematogenous dissemination mucormycosis model: Seven groups of male BALb/c mice were examined in control, infected, and treated groups with Liposomal amphotericin B. (3) Human mucormycosis: in this study, two samples of patients with rhinocerebral mucormycosis with diabetes mellitus treated and untreated with liposomal amphotericin B were examined. Total RNA extraction and cDNA synthesis were performed from the studied samples. The relative expression changes of the target genes and miRNAs were evaluated using real-time PCR carried out using Sybrgreen-based detection methods.

Results
Monocyte-derived macrophages had a steady pattern in relative changes in gene expression. An increase in expression of two genes, GRP78 and CotH3, was observed in the samples, and all miRNAs targeted by the GRP78 gene included hsa - miR-16-5p; hsa -miR-335-5p and hsa -miR-93-3p showed a decreasing pattern. In the mice mucormycosis model, relative gene expression changes were observed, and mmu-miR-181b-5p showed increased expression deviation in all groups. The clinical sample of diabetic patients with untreated rhinocerebral mucormycosis also had a consistent pattern of GRP78 and CotH3 increased gene expression. The hsa-miR-16-5p and hsa-miR-335-5p have increased expression, while hsa-miR-93-3p decreased expression.

Conclusion
After validation, these micro RNAs can be used as valuable markers in mucormycosis detection and treatment processes.