bluuebunny/crossref_metadata_2025 huggingface dataset:
P424 Comparative analysis of Galactomannan Lateral Flow Assay, Galactomannan Enzyme Immunoassay and BAL culture for diagnosis of COVID-19-associated pulmonary aspergillosis
Authors: Hedayati Mohammad , Ghazanfari Mona , Charati Jamshid Yazdani , Davoodi Lotfollah , Arastehfar Amir , Moazeni Maryam , Abastabar Mahdi , Haghani Iman , Mayahi Sabah , Hoenigl Martin , Pan Weihua
Published: September 2022

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

 

Objectives

Invasive pulmonary aspergillosis is a well-known complication in COVID-19 patients. A reliable and early diagnosis is essential for the early initiation of antifungal therapy. On the other hand, conventional Bronchoalveolar lavage (BAL) culture may lack the specificity of differentiating between colonization and infection. Here, we aimed a comparative analysis of Galactomannan Lateral Flow Assay (GM-LFA), Galactomannan Enzyme Immunoassay (GM-EIA), and BAL culture for diagnosis of COVID-19-associated pulmonary aspergillosis (CAPA).

Methods

BAL (n = 105) and serum samples (n = 101) from COVID-19 patients who were mechanically ventilated for ≥4 days in intensive care units (ICUs) were evaluated by GM-LFA and GM-EIA. All BAL samples were cultured on Sabouraud-Chloramphenicol dextrose Agar. Diagnostic performance of GM-LFA and GM-EIA in BAL (GM indexes ≥1) and serum (GM indexes >0.5) were evaluated and sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) and Roc curve compared with BAL culture calculated.

Results

The Aspergillus GM-LFA for serum and BAL samples showed a sensitivity of 46.9% and 54.5%, specificity of 100% and 91.7%, PPV of 100% and 75%, and NPV of 80.2% and 81.5%, when compared with BAL culture, respectively. GM-EIA showed sensitivities of 56.3% and 60.6%, specificities of 94.2% and 88.9%, PPVs of 81.8% and 71.4%, and NPVs of 82.3% and 83.1% for serum and BAL samples, respectively.

Conclusion

According to our results BAL GM detection using both EIA and LFA may be a promising approach for early diagnosis of CAPA and differentiating between colonization and invasive infection.
P428 Evaluation of Galactomannan enzyme assay in non-hematological non-neutropenic ICU admitted patients for diagnosis of Invasive Pulmonary Aspergillosis as per EORTC criteria 2019
Authors: Kashyap Sanchi , Capoor Malini R. , Lavanya P. , Gupta Poonam , Verma P.K. , Talwar Vandana , Sachdeva Harish Chand
Published: September 2022

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

 

Objective

The objective of this study was to evaluate galactomannan enzyme assay in non-hematological ICU patients for diagnosis of Invasive Pulmonary Aspergillosis as per EORTC criteria 2019.

Methods

Galactomannan detection in serum samples was performed by enzyme-linked immunosorbent assay in 138 patients with clinically suspected pulmonary aspergillosis between January 2012 and January 2019 in patients admitted to ICUs. Patients with hematological diseases or who underwent HSCT were excluded. Control group (n = 25) was selected from patients with no obvious immunosuppression, cough/dyspnea, or any other respiratory symptoms, etc admitted for elective surgeries from surgery wards. According to the criteria of the European Organization for Research and Treatment of Cancer/Mycoses Study Group revised in 2019, the patients were categorized as proven, probable, and possible IA. Galactomannan optical density indices ≥0.5 and ≥1 were analyzed with respect to clinical, radiological, and mycological evidence.

Results

Amongst 138 patients 2 fulfilled the criteria of proven IA, 78 were probable IA, and the rest (58) were grouped under possible IA. In the control group, the Galactomannan optical density index was ≤ 0.5. Receiver operating characteristic curve analysis showed the serum GM detection cutoff value was 0.91 (95% CI 0.885-0.973, P-value <.0001) with Youden index J = 0.78, its diagnostic value for pulmonary aspergillosis was optimized, and the sensitivity and specificity reached 83.75% and 94.83% respectively. Other cutoffs had high variance between sensitivity and specificity for the diagnosis of IPA.

Conclusion

Our study highlights the usefulness of the serum GM antigen test in the early diagnosis of IA and suggests a GMI cutoff of 0.91 as it has the highest diagnostic accuracy. Also, we recommend that patients with a GM OD of ≥1.0 should be labeled as clear GM positive, whereas those with <0.5 OD should be labeled as GM negative, and in those with OD ranging from 0.5 to 1.0, repeat sampling from the patient is advisable. It seems more reasonable not to overtreat all patients with fever refractory to broad-spectrum antibiotics with antifungal agents, but rather to decide taking into account the serum GM antigen positivity. A negative GM antigen can curtail the usage of unwarranted antifungal therapy, especially in resource-poor country like India.
P431 Tip to correct the variation position error in applying long-read high-throughput sequencing technology for fungal identification
Authors: Langsiri Nattapong , Worasilchai Navaporn , Irinyi Lazlo , Meyer Wieland , Chindamporn Ariya
Published: September 2022

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

 

 

Presently, long-read high-throughput sequencing technology has been started applying in the medical mycology field instead of common Sanger sequencing. With its capability to sequence nucleic acid among the large region compared to Sanger sequencing, this technology can detect and analyze both inter- and intra-species sequence variation in internal transcribed spacer (ITS), a fungal house-keeping region, resulting in high accurate fungal identification. However, to set up the long-read high-throughput sequencing technology for fungal identification is quite challenging because of several factors related to the accuracy including the variation position error. So, in this study, we summarize the tip to correct the variation position error in applying long-read high-throughput sequencing technology for fungal identification that we have learnt in the preliminary laboratory setting using the 8 clinical isolates of yeast: Candida albicans (n = 2), C. tropicalis (n = 1), C. glabrata (n = 1), Trichosporon asahii (n = 2), Pichia kudriavzevii (n = 1), Cryptococcus neoformans var. grubii (n = 1). Based on the recruited strains, we found that self-assembling reference sequence generated from raw data of reading by using an auto-program named Canu causes the size-inflated sequence, a larger size calculated as 22.83 ± 7.56% than it should be, resulting in the shift of variation position. This error can be corrected by the alignment process of the reference sequence with the known sequence, both size and position, prior to doing the raw read alignment. The advantage of this process could correct not only for position shifting caused by the analysis process but also the random error generated from nanopore system. To validate this correction protocol, more samples are needed for further study.
P434 Anti fungal susceptibility of Malassezia to azoles by broth microdilution method and their phylogenetic relationship based on multi-locus sequence analysis
Authors: Romald Packia Nancy , Kindo Anupma Jyoti , Veereraghavan Mahalakshmi , Umadevi U.
Published: September 2022

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

 

Objectives

Method

Part 1—Epidemiological pattern, phenotypic Identification, and anti-fungal susceptibility pattern by broth microdilution method

Part 2-Molecular Identification, Sangers Sequencing, Phylogenetic Analysis and Dendogram

Molecular Identification

Results

Figure 1 Epidemiological pattern, phenotypic Identification, and AFST by broth microdilution

Figure 2 Molecular Identification, Sangers Sequencing, Phylogenetic Analysis, and Dendrogram

Conclusions
P436 A study to compare detection of Aspergillus galactomannan in patients with clinically suspected invasive Aspergillosis using the Lateral Flow Assay (LFA) in comparison with Enzyme Linked Immunosorbent Assay (ELISA) and identification of new risk factors as per BM-AspICU criteria
Authors: Padaki Priyadarshini , Savio Jayanthi , Ramachandran Priya , Krishna Bhuvana
Published: September 2022

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

 

Objectives

Methods

A total of 50 consecutive leftover samples (30 blood and 20 BAL) received for testing by Galactomannan assay by ELISA method were included. LFA and ELISA were performed as per manufacturers’ instructions. Results were recorded using the Digital Reader provided which gives readings as Positive ≥0.5 and <0.5 Negative. Additionally, results were read visually by two different people and graded as 0 (negative), 1+ (very faint band), 2+ (faint band), and 3+ (strong band). The values obtained by ELISA were reported as Galactomannan Index. Results of LFA and ELISA were compared. Patient profiles were studied to identify the possible and probable cases of IA as per BM-AspICU guidelines.

Results

Overall, 92% samples (n = 46) were positive with ELISA. Results of LFA and ELISA were comparable in 68% samples (n = 34). For all except two samples, visual readings of LFA (5 with 3+, 12 with 2 + and 19 with 1 + readings) correlated with digital readings. Sensitivity of LFA was 73.9% (95% CI). Applying BM-AspICU criteria, 28% (n = 14) were categorized as probable, 64% (n = 32) as possible cases of IA and 8% (n = 4) as NO IA. A total of 12 patients had classical host factors, 42% (n = 21) had other risk factors and we were able to identify 16% (n = 8) cases with new risk factors which were CNS involvement/brain injury/hemorrhage on mechanical ventilation. Clinical features were present in all of these 58% (n = 29) cases whereas radiological imaging was done in only 25 cases out of which 28% (n = 14) were positive. BM-AspICU criteria require ≥2 mycological criteria, but only two of the BAL samples were culture positive for Aspergillus species. These criteria could not be strictly applied to most patients in our study since invasive procedures such as BAL was not done, instead, blood was sent for galactomannan detection.

The average turn-around time was 3-7 days for ELISA since the test was run twice a week whereas it was 5-24 h for the LFA. The pre-processing of samples and time required to perform the procedure for both the assays are similar. The cost per test for ELISA was Rs.1675/- as against LFA for Rs.750/-.

Conclusion

LFA for galactomannan detection is an easier to perform, cost-effective method for diagnosis of IA as compared with ELISA. LFA overcomes the difficulty of insufficient volume of calibrators and controls provided with the ELISA kit. Useful especially for patients admitted in ICU where reports can be issued on the same day instead of batch testing. Identification of new risk factors and inclusion of the same under guidelines is essential to suspect and send samples for testing for early initiation of treatment.
P440 General protocol applied to the identification and production of new biomarkers with potential use for the diagnosis of histoplasmosis
Authors: Arias Juan David Puerta , Isaza Juan Pablo , Moreno Ernesto , Berrio Indira , Restrepo Luz Elena Cano , Naranjo Tonny
Published: September 2022

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

 

Objective

To identify and produce novel biomarkers with potential use for the specific diagnosis of H. capsulatum infection.

Methods

Here, we design a novel strategy to search and select new Candidate genes for biomarkers that integrates the use of a computational analysis model that includes the application of bioinformatic tools such as OrthoMCL, BLASTp, TargetP, and SignalP, applied on a local collection of proteome database obtained manually from GenBank-NCBI, and the analysis of previously published biological and experimental data sets, including a secreted proteome database obtained from pathogenic yeast-phase H. capsulatum culture filtrates, a Histoplasma yeast and mycelial transcriptomes database, and a urine-peptides database from Histoplasma-immunoassay-positive patients.

For the synthesis of the Candidates, an internal protocol for the production of recombinant proteins in prokaryotic and eukaryotic systems was applied. Obtaining polyclonal antibodies (PAb) specific for each biomarker was carried out by adapting a rapid immunization protocol for BALB/c mice.

Finally, the computational model was experimentally validated, evaluating the reactivity and specificity of PAb anti-Histoplasma with fungus culture extracts and samples from patients with histoplasmosis.

Results

Using the computational analysis model, 2 Candidate genes for diagnostic biomarkers were identified. Subsequently, the construction of expression vector for each Candidate and the production of these genes were achieved using a standardized protocol for the production of recombinant proteins.

Polyclonal antibodies (PAb) anti-histoplasma were obtained and shown to be reactive against purified H. capsulatum-antigens. Finally, we confirmed the presence of these antigens in yeast culture extracts of H. capsulatum and demonstrated the immunoreactivity of anti-Histoplasma PAb with urine samples from patients previously diagnosed with histoplasmosis.

Conclusion

The generation of novel strategies that combine data analysis, computational tools, and transcriptomic and proteomic techniques could be very useful for the identification of new biomarker genes and the development of microbiological diagnostic tests for important pathogens.
P444 Detection of causative agents of infectious keratitis in patients from western rajasthan
Authors: Shrimali Twishi , Sharma Anuradha , Ghosh Anup , Morya Arvind , Tak Vibhor , Nag Vijayalakshmi
Published: September 2022

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

 

Objective

To determine the spectrum of causative agents, the related risk factors, and their association in patients of infectious keratitis.

Methodology

It was a prospective study conducted over a period of 18 months from August 2018 to January 2020, which included 100 patients attending the Ophthalmology OPD with features of keratitis. Ophthalmological examination was followed by corneal scrapings’ collection, which were subjected to culture, microscopy, and molecular diagnostic tools. Bacterial isolates were identified by conventional methods and MicroScan Walkaway system while the isolated fungi were identified conventionally. Pan-fungal primers were used to detect fungal elements directly from the sample.

Results

Out of 100, 41 cases were positive by culture, of which 32 (78.04%) had fungal and nine (21.95%) had bacterial keratitis. Fusarium spp. accounted for 33.33% of fungal and Pseudomonas aeruginosa accounted for 55.55% of the bacterial isolates. Fungal material was detected in 41% using pan fungal primers. Cases were maximally recorded during July-October. Traumatic history was present in 78% patients caused by vegetative matter (49%). A male preponderance (67%) was also observed. Four patients underwent evisceration in spite-of rigorous management.

Conclusion

Poor prognosis emphasizes the need for faster diagnostics, which can detect the causative agents from the clinical specimen itself, reinforcing the concept of clinical metagenomics.
P445 Clinical evaluation of the performance of the most commonly used eumycetoma diagnostic tests using sequencing of the internally transcribed spacer region as the golden standard
Authors: Siddig Emmanuel , Nyuykonge Bertrand , Mhmoud Najwa , Abdallah Omnia , Bahar Mustafa , Ahmed Eiman , Nyaoke Borna , Zijlstra Eduard , Verbon Annelies , Bakhiet Sahar , Fahal Ahmed , van de Sande Wendy
Published: September 2022

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

 

Objective

Mycetoma is a neglected tropical skin disease, caused by 70 different causative agents. For most of the causative agents, molecular identification is the only reliable method to identify the species level. In practice, ultrasound, histopathology, culturing, and species-specific PCRs are most commonly used for species identification. However, the performance of these different tests was not validated using molecular identification by sequencing barcoding genes.

Methods

In this study, we validated the performance of the most commonly used diagnostic tools including culture, histopathology, Ultrasound and two species-specific PCR for Madurella mycetomatis on 222 patients suspected of fungal mycetoma by M. mycetomatis; the sensitivity, specificity, and accuracy of each method was calculated.

Results

From the 222 patients, 154 (69.3%) were correctly identified by ultrasound, histology, culture, and both species-specific PCRs. For five patients all tests were negative and for three only the ultrasound was indicative of mycetoma. For the other 60 patients, at least one of the assays was negative for M. mycetomatis. The two species-specific PCRs were the most sensitive and specific, followed by culture and histology. Ultrasound was the least specific as it only allows to differentiate between actinomycetoma and eumycetoma. However, with ultrasound, an identification could be obtained in 9.38 min. PCR took 3.76 h, histology 8.5 days, and culturing 21 days.

Conclusion

We concluded that PCR directly on DNA isolated from grains is the most rapid and reliable diagnostic tool to identify M. mycetomatis from eumycetoma grains to use species-specific PCRs. In order to shorten the time to identification of other causative agents, the focus should be on developing more molecular assays for those species.
P457 Clinical correlation of beta galactomannan with culture and patient outcome in a tertiary care center in south india
Authors: K. Vichitra , Kindo Anupma
Published: September 2022

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

 

Background

Invasive aspergillosis has increased in the last decade. They mainly occur in neutropenic patients (following anticancer treatment) and in patients treated with immunosuppressants and corticosteroids. The galactomannan antigen in serum appears to be a serological method able to aid in the diagnosis of invasive aspergillosis also the antigen detection in bronchoalveolar lavage has proven to be advantageous for the diagnosis of invasive aspergillosis.

Objectives

Methods

A total of 175 samples were collected from patients suspected to have fungal infections from the period of January 2018 to March 2022 from a tertiary care center in South India. The galactomannan assay and culture were done for these samples. Data of age, sex, gender, diagnosis, underlying conditions, antifungal treatment, and outcomes were collected.

Results

Out of 175 samples collected from patients suspected with Aspergillosis, 120 were males and 55 were females (7 were repeat samples).

The major underlying conditions were diabetes and hypertension, coronary artery disease (CAD), and Hematopoetic malignancy such as B Cell lymphomas, ALL, and AML (Fig. 1).

Conclusion

Galactomannan alone cannot be taken as a diagnostic marker. Clinical correlation, radiologic findings, as well as underlying risk factors play a major role for decision on the initiation of empiric treatment.
P465 Aspergillosis in free-ranging Magellanic penguins
Authors: Trápaga Mariana , Melo Aryse Martins , Poester Vanice , Canabarro Paula Lima , Silva Andrine , Estima Sérgio , Frere Esteban , Xavier Melissa Orzechowski
Published: September 2022

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

Aspergillosis is an opportunistic fungal disease caused by the Aspergillus genus, mostly by Aspergillus section Fumigati. Captive Magellanic penguins are vulnerable to Aspergillus infection, being this mycosis a limiting factor during the process of rehabilitation with a mortality rate of around 50%.

Objectives

Giving the scarce data regarding the occurrence of aspergillosis in non-captive penguins, we aimed to evaluate the proportional mortality by aspergillosis in free-ranging Magellanic penguins during their migration and reproductive season.

Methods

Carcasses of Magellanic penguins were collected from the Southern coast of Brazil between June 2017 and October 2019 between Barra do Chui beach (Southern RS, Brazil - 33°44'19.9‘S 53°21'56.3’W) and the isthmus with the Lagoa do Peixe (Southern RS, Brazil - 31°26' S, 51°10' W 31°14'S, 50°54'W). In addition, in January 2019, penguins found dead in the reproducing colony in four islands localized in Puerto Deseado City (Santa Cruz, Patagonian, Argentina - 47°45′00″S 65°55′00″W) were collected. All animals were necropsied, and macroscopic alterations were observed. Samples of macroscopic lesions and/or respiratory systems were collected for histopathology and mycological culture. Only proven aspergillosis cases, defined by suggestive lesions at necropsy, associated with hyaline, septate, and 45° branched hyphae in histopathological slides and isolation of Aspergillus sp. in the culture were computed. Fungal isolates were identified by molecular techniques.

Results

A total of 98 Magellanic penguins were included in our study, being 80 recovered on the Southern RS beach, and 18 from the Patagonian colony. Two penguins collected in Southern Brazil were diagnosed with aspergillosis, both juveniles, one showing nodules in the lung parenchyma, and the other nodules and fungal colonies at the lung and air sac, resulting in a proportionate mortality rate of 2.5%. Both isolates were identified as A. fumigatus sensu stricto. Regarding the carcasses collected from the reproductive colony (Patagonian islands), no penguin had anatomopathological or mycological evidence of aspergillosis.

Conclusion

Given the already known importance of aspergillosis in seabirds undergoing rehabilitation, these data suggest that penguins may already arrive in these centers infected by Aspergillus spp. The absence of aspergillosis cases in the reproductive period could be attributed to the low number of carcasses included. Our study is an initial step to demonstrate aspergillosis as one of the causes of mortality also in free-living penguins, especially during the migration process, instigating more studies, in other routes of migration, as well as in reproductive colonies.
P467 Aspergillosis in Humboldt penguins — susceptibility patterns of clinical and environmental isolates from a Belgian zoo, 2018-2022
Authors: Debergh Hanne , Goens Karine , Vercammen Francis , Saegerman Claude , Packeu Ann
Published: September 2022

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

 

Objectives

Avian aspergillosis causes a heavy burden on birds in captivity, such as Humboldt penguins. In recent years the colony of a Belgian zoo has experienced very high mortality rates and the zoo has already taken several measures to lower the burden. This study was set up to see if the penguins acquire A. fumigatus via the environment and if so, if additional measures can be taken to limit the incidence.

Methods

A total of 29 clinical strains collected from 2018 to 2022 were included in the study. From April 2021 until January 2022, four samplings have been performed, accounting for every season. A combination of sand, water, nest swabs, and air was analyzed for the presence of (azole-resistant) A. fumigatus. In brief, air samples were collected at fixed locations in the penguin enclosure by impacting 1000 L on an agar plate [malt + chloramphenicol (MC) and MC + tebuconazole (MC + T)]. A total of 100 mL water was collected and 50 mL water was filtered through a 0.22 μm filter and placed on an MC plate, and repeated with the other 50 ml on MC + T. A total of 9 ml 0.1% Tween 20 + 0.85% NaCl was added to 1 g of sand and vortexed for 1 min. Both MC and MC + T plates were incubated with 100 μL of this suspension. All plates were incubated at 48°C ± 1°C for 48 h. The phenotypical resistance pattern of all clinical isolates was determined using the EUCAST method.

Results

The phenotypical resistance pattern showed resistance in 7 isolates (24%) with 5 of them showing resistance to posaconazole, one was resistant against voriconazole, isavuconazole, and posaconazole, and 1 showing pan-resistance. No A. fumigatus colonies could be detected from water samples, nor from the sand. One A. fumigatus isolate was retrieved from the nest swabs. In total 64 A. fumigatus colonies were isolated from air samples collected on MC + T medium. All have been subjected to EUCAST microbroth dilution determination and 4 resistant isolates could be detected (all had a MIC value for posaconazole of 0.5 μg/ml and one strain showed additional resistance against isavuconazole with a MIC of 4 μg/ml. Cyp51a sequencing of all resistant strains is ongoing and will give more insight in the molecular mechanisms involved to investigate the potential link with the environment.

Conclusion

This study showed high resistance rates in the clinical isolates. Four resistant isolates were found in environmental air samples. Sequencing of the cyp51A gene will give more information on a possible relation between the resistance mechanisms found in the clinical and the environmental isolates. More research should be done to investigate the origin of the resistant isolates in the environment.
P468 Aspergillosis in a colony of Humboldt penguins (Spheniscus humboldti) under managed care: a clinical and environmental investigation in a French zoological park
Authors: Perraud-Cateau Estelle , Leclerc Antoine , Cartier Noémie , Valsechi Isabel , Bailly Eric , Senechal Ronan Le , Becerra Margaux , Gallou Brice Le , Lavergne Rose-Anne , Chesnay Adélaïde , Robin Jean-Patrice , Cray Carolyn , Goddard Nicolas , Thorel Milan , Guillot Jacques , Mulot Baptiste , Desoubeaux Guillaume
Published: September 2022

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

 

Objectives

Aspergillosis is an opportunistic fungal infection due to Aspergillus spp., and primarily A. fumigatus. The disease is pervasive in avian populations, especially those under managed care. The incidence and the management of avian aspergillosis can be critically impacted by exposure to high levels of spores via environmental contamination and by strains resistant to azole drugs. The present study aimed at assessing the impact of environmental contamination on the clinical incidence of aspergillosis in penguins managed under human care. ZooParc de Beauval was chosen for the investigations, because this zoological facility hosts a large colony of Humboldt penguin (Spheniscus humboldti) and it is geographically located in the rural countryside of the Loir-et-Cher district, France, surrounded by numerous large crop fields

Material and methods

The fungal contamination in the environment was assessed through three serial sessions of surface sampling in bird nests: all isolates were counted, identified by DNA sequencing, and then systematically screened for resistance mutations and MICs (minimal inhibitory concentrations) elevation for the A. fumigatus strains. In addition, the clinical incidence of aspergillosis was evaluated in the penguin population over a 3-year period. A microsatellite-based analysis tracked A. fumigatus isolates circulation.

Results

Environmental investigations highlighted a substantial increase in the fungal load during the Autumn season (>12-fold vs. the other timepoints) and a large overrepresentation of species belonging to the Aspergillus section Fumigati (ranging from 22.7 to 94.6%) (Fig. 1). Only one cryptic species (A. nishimurae) and one isolate exhibiting resistance mutation (G138S in the cyp51A gene; MIC itraconazole >4 μg/ml) were detected. The overall incidence of aspergillosis was measured at ∼3.4% case-years and was observed mostly in juvenile penguins (Fig. 1). The analysis of microsatellite polymorphism revealed a high level of genetic diversity among A. fumigatus isolates, except for one strain that was largely over-represented during the Autumn sampling session. A limited number of isolate genotypes was collected from clinical cases and from the environment.

Conclusions

Fungal environmental contamination and incidence of aspergillosis in penguins appeared variable depending on seasons, but the rural location of the penguin habitat did not seem to influence the emergence of resistant strains.
P471 Feline sporotrichosis outcome and its impact in public health in Southern Brazil
Authors: Esperon Bruna , Poester Vanice , Munhoz Lívia , Trápaga Mariana , Sanchotene Karine , Benelli Jéssica , Klafke Gabriel , Xavier Melissa
Published: September 2022

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

Sporotrichosis due to Sporothrix brasiliensis is an emerging and neglected disease in Brazil. Domestic cats are susceptible to a severe presentation of this mycosis, carrying a high fungal load in their lesions. They frequently infect other animals and even humans by scratches and/or bites. Thus, the correct management and treatment of feline sporotrichosis are crucial aspects of the control of the disease in a population.

Objective

We aimed to evaluate the management and outcome of feline sporotrichosis cases in a hyperendemic city in southern Brazil (Rio Grande do Sul state).

Methods

Database from the Mycology Laboratory (LabMyco) of the Federal University of Rio Grande (FaMed–FURG) was consulted to gather data from all proven feline sporotrichosis cases (confirmed by mycological culture), between January 2019 and December 2021. It was included in this study in all cases in which the phone number of the cat owners was available. All of them were contacted and invited for an interview by quick and short questions regarding the management and the outcome of their cats with sporotrichosis. Disagreement to participate, and change/incorrect phone number contacted were used as exclusion criteria.

Results

During the 3-year period studied a total of 62 owners, from 165 felines diagnosed with sporotrichosis in the LabMyco, had a phone number available. A total of 35 owners were excluded, totalizing 27 participants in this study. More than half (51.8%; n = 14) reported returning only once to the veterinarian to clinical accomplishment, 48.1% (n = 13) of them do not use personal protective equipment to handle the infected animal, 44.4% (n = 12) highlighted the difficulty in daily administering drugs to the cat and only 18.5% (n = 5) affirmed to had isolated the infected animal during the treatment. Two animals (7.4%) with advanced signs of disseminate sporotrichosis died before starting treatment, and the others received itraconazole and/or potassium iodide as the drug of choice. Clinical cure was achieved in 40% of the cats treated (10/25), 28% (7/25) evolved to death, 16% (4/25) are still in treatment due to new lesions (recidive), and the other four animals were abandoned in the streets. In addition, zoonotic transmission occurred in three (11.1%) owners, which developed lymphocutaneous sporotrichosis after a scratch or bite by the infected cat.

Conclusion

Sporotrichosis in Southern Brazil is a public health threat, in view of this, our study shows the urgent necessity of government strategies and interventions that promote health education and implement a service to attend, and provide treatment accomplishment to feline sporotrichosis in view of control the current hyperendemic of this mycosis.
P472 The mechanism of action of antifungal activity of Zanthoxylum armatum fruit's oil against Candida cells does not involve ROS generation
Authors: Pasrija Ritu
Published: September 2022

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

 

Objective

To explore the antifungal activity of Zanthoxylum armatum fruit's oil against different Candida species and its mechanism of action.

Methods

The Z. armatum fruit's oil activity was assessed against C. albicans, C. krusei, C. glabrata, C. parapsilosis, C. tropicalis, and C. guilliermondii through different drug susceptibility assays including, MIC, agar diffusion and spot assay. The mechanism of action was explored through sterol analysis, germ tube inhibition, epithelial cells adherence, and ROS generation.

Results

The oil from the fruits of Z. armatum was subjected to GC-MS analysis, and linalool (72%) was found as the major component. The drug susceptibility measured through different methods, including minimum inhibitory concentration (MIC), where end-point was 3% v/v for different species tested, and the same pattern was observed in agar diffusion and spot assay. The antifungal activity was found to be fungicidal in nature and the major reason appeared to be the reduction in ergosterol levels inside cells. It resulted in lowered germ tube formation, an important indicator in virulence of C. albicans. The oil reduced adherence of Candida cells to buccal epithelium significantly, which is the first step in invasion, biofilm formation, and damage to oral epithelial cells. Interestingly, unlike most antifungals, where reactive oxygen species generation mediated killing is involved, was not found significant in the present study.

Conclusions

The Z. armatum fruits oil exerts its antifungal activity by inhibiting ergosterol formation and reduced germ tube formation.
P481 Antifungal vaccines: An alternative approach to combat multidrug resistance in the post-antibiotic era
Authors: Ahmad Aijaz
Published: September 2022

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

Alexander Fleming's discovery of the antibiotic ‘Penicillin’ and many more has inspired after saving millions of lives and spared immeasurable suffering worldwide. However, he also warned about the consequences of abuse of antibiotics that could lead to the expiry of the golden age of antibiotics. Current decreasing effectiveness of antibiotics in general and antifungals in particular, evolution of new drug-resistant forms, and species and lack of commercial interests to develop new antifungals is a clear indication of a post-antibiotic era. There is an amplified warning for the pharmaceutical industries, academic and research institutions, and governments to invest the necessary resources to produce next-generation, safe and effective antifungal vaccines. Candida auris is one of the emerging pan-resistant pathogenic yeast among immunocompromised patients worldwide. Early diagnostics and vaccines are the best alternatives to handle this drug-resistant pathogen. Cell wall proteins are responsible for eliciting an immune response in the host and therefore, can be promising candidates for the development of therapeutic biomarkers and vaccines against C. auris.

 

Antifungal susceptibility profiling against standard antifungal drugs was performed using CLSI guidelines. The cell wall proteome was characterized by using LC-MS followed by bioinformatic and functional analyses. Furthermore, the protective effect of the cell wall-associated protein against C. auris infection was evaluated in BALB/c female mice.

 

A total of 60 proteins were predicted to be immunodominant. In vivo results demonstrated lowered fungal burden in target organs and increased survival in vaccinated mice as compared with the infection control group revealing the immunogenic property of cell wall-associated proteins.

 

This study validates the mass-spectrometry approach to identify antigenic proteins of C. auris and the potential of these proteins to serve as biomarkers for the development of diagnostic assays and/or vaccines against infections caused by C. auris.
P482 Burden of serious fungal infections in India
Authors: Ray Animesh , K Adarsh Aayilliath , Banerjee Sayantan , Chakrabarti Arunaloke , Denning David W.
Published: September 2022

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

 

 

Fungal disease is frequent in India but its incidence and prevalence are unclear. Literature searches yielded over 2900 papers; 434 papers with incidence/prevalence/proportion data were analyzed. Deterministic modeling allowed annual incident and prevalence estimates for multiple life and sight-threatening infections with significant morbidity. An estimated 55 916 148 of 1 393 400 000 people in India (4.01/100 000) suffer from a serious fungal disease. The prevalence (in millions) of recurrent vulvovaginal candidiasis is 24.3, allergic bronchopulmonary aspergillosis (ABPA) is 2.0, tinea capitis in school-age children is 25, severe asthma with fungal sensitization of 1.36, chronic pulmonary aspergillosis of 1.75 and chronic fungal rhinosinusitis of 1.52. The annual incidence of Pneumocystis pneumonia (58 400), invasive aspergillosis (250 900), mucormycosis (195 000), esophageal candidiasis in HIV (266 600), candidemia (18 800), fungal keratitis (1 017 100) and cryptococcal meningitis (11 300) were also determined. Histoplasmosis, talaromycosis, mycetoma, and chromoblastomycosis are less frequent. India's fungal burden is high and under-appreciated in clinical practice.

See Figures below.
P483 Invasive fungal infection in hematopoietic stem cell transplant recipient from an Indian oncology setting
Authors: Ghafur Abdul , Das Bikram , Karthik Radhika , Easaw Benjamin M , Raja Ramanan T , Easaw Jose
Published: September 2022

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

 

Objectives

Invasive fungal infections (IFI) are one of the major causes of morbidity and mortality in post-hematopoietic stem cell transplant (HSCT) recipients. Data from India are limited. The objective was to analyze the incidence, risk factors, and outcomes associated with IFI in our center.

Methods

Adult patients, who underwent marrow/stem cell transplantation between 2014-2018, in an oncology center in India, were included in this retrospective observational study. The revised consensus definition of IFI by the European Organization for Research and Treatment of Cancer/Invasive Fungal Infections Cooperative Group and the National Institute of Allergy and Infectious Diseases Mycoses Study Group (EORTC/MSG) in 2008, was considered to define cases. Incidence, risk factors, and outcomes associated with IFI were analyzed.

Results

Out of the 126 patients who underwent HSCT between 2014-2018, 56 (44.4%) had Allo HSCT, 64 (50.8%) had auto HSCT and 6 (4.8%) had haplo-identical HSCT. A total of 83 (63%) were males and 43 (34%) females, 113 (83.9%) Asians, and 13 (10.3%) Africans. Total 111 (88%) patients received myeloablative conditioning and 24 (19%) received total body irradiation. The hematological conditions were acute myeloid leukemia (AML) n = 23 (18.25%), acute lymphoblastic leukemia (ALL) n = 16 (12.69%), chronic myeloid leukemia (CML) n = 4 (3.17%), Hodgkins lymphoma (HL) n = 17 (13.4%), non-Hodgkins lymphoma (NHL) n = 11 (8.73%), Myeloma n = 35 (27.7%), sickle cell disease n = 13 (10.31%), etc. Most patients received fluconazole 78 (61.9%) followed by micafungin 23 (18.25%), posaconazole 20 (15.87%), voriconazole 4 (3.17%), and liposomal amphoterin B 1 (0.79%) as antifungal prophylaxis. The overall rate of IFI (possible cases included) was auto-HSCT n = 5 (7.81%), and Allo-HSCT n = 5 (8.92%). Among auto-HSCT, the IFI was Proven = 0, Probable n = 1 (1.5%), and Possible n = 4 (6.25%), and among Allo-HSCT Proven = 0, Probable n = 2 (3.57%), and Possible n = 3 (5.35%). These cases had IFI lung based on imaging and serological tests. None of the cases had a lung biopsy. There were no incidents of candidemia. No patients in Haplo-HSCT had IFI. The 1-year survival rate among the IFI cases was 8/10 (80%). As the number of patients with IFI was very low, a meaningful comparison of the risk factors, and the impact of prophylactic regimens were difficult.

Conclusions

The overall rate of IFI in HSCT patients in our setting was low compared to global data.
P496 Effect of fluconazole prophylaxis in prevention of local fungal infections in patients of head and neck cancers receiving chemo-radiotherapy
Authors: Lohia Shashwat , Suri Kapil , Capoor Malini
Published: September 2022

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

 

Objectives

To ascertain, in head and neck cancer patients receiving chemo-radiotherapy, the efficacy of fluconazole prophylaxis in

1. Reducing the incidence of oropharyngeal fungal infections

2. Reducing the incidence of severe i.e., grade 3 and 4 oropharyngeal mucositis

Methods

A total of 66 cases of Stage III or IV squamous cell carcinoma of the oral cavity or oropharynx who were candidates for radical chemo-radiotherapy (CCRT) were randomly assigned between arm A (study arm) and arm B (control arm) using computer-generated algorithms. Both arms received conventionally fractionated radiation (66-70 Gy) with concurrent weekly intravenous cisplatin (40 mg/m2) along with standard measures of oropharyngeal hygiene. In addition, arm A received Tab Fluconazole 100 mg/d administered per oral after lunch from initiation to the completion of radiotherapy, while arm B received Tab Fluconazole 100 mg/d, if fungal infections appeared, for a period of 7-14 days.

Weekly clinical examination for oro-pharyngeal candidiasis was done. Throat swabs for microbiological assessment of fungal colonization were done before treatment (zero week), during second and sixth week of CCRT, and 4 weeks after the completion of CCRT, and if any clinical suspicion of fungal infection was present.

Results

See Figures below.

Conclusion

Tab fluconazole 100 mg prophylaxis in head and neck cancer patients receiving chemo-radiation is
P500 Validation panel for MALDI-TOF identification of fungi
Authors: Stubbe Dirk , Becker Pierre , D'hooge Elizabet , Debergh Hanne , Packeu Ann
Published: September 2022

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

Sciensano, the Belgian federal scientific institute for public and animal health, houses the BCCM/IHEM Fungi Collection which contains more than 15 000 strains, belonging to over 1 500 different species. The collection is managed according to ISO 9001 standards.

 

Its purpose is to make fungal strains available for academics, clinicians, industry, and education.

 

Fungal pathogens are not as often encountered as bacteria in the clinical laboratory. Additionally, laboratories may not have the knowledge or logistics for the long-term preservation of axenic fungal isolates. Without an array of fungal strains with confirmed identity, it is complicated to implement new protocols and equipment when these need to be validated for the identification of fungi.

 

To short-cut this problem and support laboratories in identifying clinical fungi in routine activities, BCCM/IHEM has developed two validation panels for the identification of fungi via MALDI-TOF mass spectrometry: there is a validation panel with yeasts and a validation panel with filamentous fungi. The selection of strains is based on species that are routinely encountered in a clinical laboratory, and also contains some rarer, but emerging fungal pathogens, like Trichophyton indotineae and Candida auris. The identity and purity of the strains in these panels have been verified according to ISO 17025 accredited protocols. This allows the laboratory to evaluate in a short term the extraction protocol, the MALDI-TOF machine, and the database of reference mass spectra.
S1.3a Genetics andgenomics of Malassezia species
Authors: Laniri Giuseppe
Published: September 2022

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

 

 

Malassezia includes yeasts belonging to the subphylum Ustilaginomycotina within the Basidiomycota. Malassezia yeasts are attracting the interest of both basic and applied scientists for their unique biological features, and for their importance in clinical and cosmetic settings. Although Malassezia yeasts are commonly found as commensal on human and animal skin, they are also associated with several skin disorders, such as dandruff/seborrheic dermatitis, atopic eczema, pityriasis versicolor, and folliculitis. More recently, an association of Malassezia with Crohn's disease, pancreatic ductal adenocarcinoma, and psoriasis exacerbation has been reported. To understand the genetic basis of Malassezia commensalism and pathogenicity, the availability of genomic and molecular tools plays a crucial role. Genomics advances in Malassezia reveal karyotype variations and gene turnover events, including genes horizontally transferred from bacteria. Moreover, the increasing availability of transcriptomic data allows us to prioritize studies on novel key genes that potentially characterize the pathophysiology of Malassezia fungi. For gene function studies, protocols for Agrobacterium tumefaciens-mediated transformation were developed and utilized in strategies of random insertional mutagenesis or targeted gene replacement through CRISPR/Cas9. Developed tools can be combined with the use of host-pathogen interaction models, such as the easy-to-use wax moth larvae of Galleria mellonella or the more complex murine skin model, enabling the characterization of both the fungal components that trigger skin damage and inflammation, and the inflammatory and antifungal response of the host to prevent fungal infection through immunological and molecular analyses of experimentally infected tissue.
S1.3c Diversity and hybridization in Malassezia furfur
Authors: Theelen Bart
Published: September 2022

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

 

 

The Basidiomycetous yeast Malassezia is the most abundant fungal genus on healthy human skin but may also cause various skin disorders such as seborrheic dermatitis, dandruff, and pityriasis versicolor. In recent years, Malassezia has increasingly been implicated in health and disease beyond the skin: as an underestimated cause of Malassezia bloodstream infections (BSIs) in immunocompromised patients and neonates, associated with Crohn's disease, promoting pancreatic oncogenesis, and exacerbating cystic fibrosis. Malassezia furfur is the number one Malassezia BSI cause and is also implicated in many skin disorders. With these new discoveries of Malassezia’s impact on human health, the need for a better understanding of its evolution and pathobiology also became more pressing. Hybridization has been suggested as a biological mechanism of adaptation to new hosts, and may lead to increased pathogenicity. Many examples of major hybrid yeast pathogens exist, such as Candida albicans, C. orthopsilosis, C. metapsilosis, and multiple examples in the Cryptococcus gattii/Cryptococcus neoformans species complex. Here the multiple hybridization events of the Malassezia furfur species complex will be discussed. Two distinct hybridization events occurred between the same parental lineages, and these parental strains were originally also hybrids. The identification of a pseudobipolar mating system and the analysis of the mating-type loci provide evidence that sexual liaisons of mating compatible cells from these parental lineages led to a diploid/aneuploid state in the hybrid lineages. Sequence similarity percentages suggest that both parental lineages in fact are two different species. The genetic diversity of ca 300 strains belonging to this species complex is evaluated in relationship to host background and phenotype.