bluuebunny/crossref_metadata_2025 huggingface dataset:
P307 Clinical profile of fusarium infections: case series from a tertiary care hospital
Authors: Vineeth VK , Sethuraman Nandini , Gopalakrishnan Ram
Published: September 2022

Poster session 2, September 22, 2022, 12:30 PM - 1:30 PM

 

Objective

To study the clinical profile of patients with Fusarium infections diagnosed at our tertiary care center during the study period from February 2017 to March 2022.

Methods

We conducted a retrospective case study wherein all consecutive patients with Fusarium infections between February 2017 to March 2022 were accessed. The diagnosis was categorized based on either fungal culture alone or fungal culture with histopathology findings.

Results

A total of 12 patients with Fusarium infections were encountered during this period. The mean age was 49. In all, 5 were females and 7 were males, and 5 patients had diabetes as a risk factor. Other risk factors included were chemotherapy for multiple myeloma (1) and lymphoma (1), polytrauma (1), and surgery after pituitary macroadenoma. A total of 8 (66.7%) patients were on antifungal prophylaxis at the time of diagnosis.

A total of 6 (50%) had localized infections whereas, remainder 6 (50%) had disseminated infection, and 25% presented with onychomycosis. Seven patients were diagnosed based on fungal culture and five were diagnosed based on histopathology findings collaborated with a fungal culture. There was one patient with Fusarium blood stream infection, who expired within 2 weeks of hospitalization.

A total of 10 patients had F. solani whereas, 2 had F. oxysporum isolated in fungal cultures. In all, 42% of patients in the study had high Beta- D-Glucan (BDG) and 67% of the patients underwent source control of the involved region. A total of 9 patients (75%) received voriconazole as antifungal treatment and 3 patients received Amphotericin B. Four patients expired, three were lost to follow up and five did not develop relapse on follow-up.

Conclusion

Fusarium is an opportunistic human pathogen severely affecting immunocompromised patients, especially patients with hematological malignancies, prolonged neutropenia, and post-hematopoietic stem cell transplantation. Our study records a notable number of Fusarium infections among diabetics and onychomycosis was a common presentation. A high index of suspicion is of utmost importance in patients with risk factors and serum BDG may help in suspicion of invasive Fusarium infections. The 33% mortality in our study stresses the need for early diagnosis and treatment.
P313 A retrospective analysis of COVID-19 - associated Mucormycosis in a tertiary care center in Northern India
Authors: Yadav Sukriti , Bora Ishani , Mohi Gursimran Kaur , Shivaprakash M R , Singh Mini P , Ratho R K , Muralidharan Manjul
Published: September 2022

Poster session 2, September 22, 2022, 12:30 PM - 1:30 PM

 

Objectives

Methods

This is a retrospective observational study with (n = 433) samples. Nasopharyngeal and oropharyngeal swabs were collected from clinically suspected cases of mucormycosis. These were then subjected to Xpert® Xpress SARS-CoV-2 test. Those samples which tested positive were included in this study. A history sheet form was designed and filled up for 238 patients who were admitted and could be traced properly.

Results

Total number of SARS CoV-2 positive samples was 296 out of 433 samples. Among the tested 66% (287/433) were male and 34% (146/433) were female. Relative positivity was higher among women 74% (107/144) than men 66% (189/285). Amongst 238 admitted patients, 83.1% patients (198/238) completed treatment and were discharged while 13.9% (33/238) succumbed during management. Diabetes was found to be most common risk factor at 88.65% (211/238) followed by hypertension 36.5% (87/238), and corticosteroid use 26.89% (64/238). Type 2 DM was the most frequent with 96% (203/211) and only 4% (8/211) patients having Type 1 DM. Amongst the diabetics, 74.8% (158/211) were on medication (oral hypoglycemic agent/insulin/both), and 16.6% (35/158) of them were non-compliant to treatment. Most common clinical presentation was facial swelling 66.8% (159/238) followed by facial pain 52.52% (125/238) and headache 34.45% (82/238). Microscopy findings either nasal or palatal scraping was positive in 69.75% (166/238) cases with aseptate hyphae in 95% (157/166) patients while septate hyphae were seen in 3% (5/166). Both aseptate and septate hyphae were present in 2% (4/166 patients).

Amongst the deceased patients 91% (30/33) had diabetes mellitus exclusively of Type 2 and among them, 33% (10/30) were not on any anti-diabetic medication. It was also observed that among the deceased people who were on anti-diabetic medication, 45% (9/20) were non-compliant to the treatment. Combinedly 63.33% (19/30) diabetics were on inadequate blood sugar level control which might be strongly contributing to mortality.

Conclusion

Percentage of clinically suspected mucor patients who were COVID-19 positive was 67%. Although male patients were twice more tested than females but females were proportionately more positive for SARS-CoV2. Type 2 diabetes mellitus was the lead risk factor, with facial swelling being the predominant presentation. Interestingly, fungal smear was positive in 166 out of 238 patients. Despite the best management, the mortality rate was 13.86% in such patients. It was also seen that untreated or non-compliance to anti-diabetic treatment resulted in increased mortality. Hence, it's necessary to review cases of COVID-19-associated mucormycosis for prompt management and to reduce morbidities and mortality.
P377 Study of the prevalence of candidemia and its antifungal susceptibility pattern in a tertiary care hospital
Authors: Sharma Alisha , Patel Ankita , Mahajan Pooja , Bahal Ashish , Ninawe Sandeep , Bhatt Puneet , Grover Naveen
Published: September 2022

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

 

Objectives

Study of the prevalence of candidemia and its resistance pattern in a tertiary care hospital.

Methods

Results

Conclusion

Candida spp. are a part of the normal flora of healthy hosts but are also found to be a major cause of invasive fungal infections which is now found to be one of the leading causes of mortality in hospitalized patients. The emergence of unusual and relatively unknown Candida species as nosocomial pathogens with increasing treatment failure, emphasizes the need to isolate and identify all species and to start early definitive treatment according to the susceptibility pattern reported to decrease mortality and morbidity rates.
P379 Antifungal activity of α defensins 5 like peptide against Cryptococcus neoformans var. grubii and in silico designing of its peptidomimetics
Authors: Singh Karuna , Neelabh  , Uttam Gunjan
Published: September 2022

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

 

Objectives

Combinatorial and retroviral therapies have been found effective in the treatment of cryptococcosis, but safer and more potent antifungal therapeutics are the need of the hour. In this regard, the natural antifungal peptides (AFPs), the essential components of the innate immune system have drawn the attention of researchers and drug developers. AFPs are 12-50 amino-acids-long host defense peptides of cationic nature. Although AFPs have certain limitations like low stability, susceptibility to proteolysis, loss of activity in the presence of metal ions and salts, short half-lives, and peptide aggregation in oral formulation, but these can be addressed by designing and synthesizing the peptidomimetics.

Methodology

The whole blood of the rat was collected by cardiac puncture. The blood was mixed with 0.83% aqueous ammonium chloride. The neutrophils were separated from lysed WBCs by centrifugation. The purification of the peptide was carried out by SDS-PAGE, size exclusion, cation exchange chromatography, and HRLCMS. The MSA of the characterized peptide was performed using a T-coffee program. The in vitro antifungal activity of crude was tested against C. n. grubii (ATCC 6352) using flow cytometry. The structure of peptides was altered through ChemDraw by replacing the elements with their bioisosteres to form mimetics. The peptide and its mimetics were docked upon the target of C. n. grubii, i.e., O-acetyl transferase (modeled through Phyre2 web server) using PatchDock, and FireDock web servers.

Results

The peptide isolated from rat PMNs had a molecular weight of 3 kDa. It has lysine and arginine which are the characteristic residues of an antifungal peptide. The eluted peptide has the highest (83%) similarity with α defensins 5 (accession no NP_001013071 XP_214 386). The mild antifungal activity of peptide was evident against C. n. grubii. The structure of O-acetyl transferase was modeled with 100% confidence and 92% query coverage (Table 1). The mimetic 1 (M1) showed the best interaction with O-acetyl transferase with −54.47 Kcal/mol binding energy and 4 hydrogen bonds (Table 2).

Conclusion

The designed M1 of α defensins 5 like peptide could be a promising lead in antifungal drug development. As mimetic 1 was designed using an in-silico approach, its synthesis and wet lab validation are desirable.
P393 Clinical spectrum and molecular characterization of Rhytidhysteron spp. (now Gloniopsis) from India
Authors: Ahmad Haseen , Kaur Harsimran , Gupta Parakriti , Shankaranarayana Shamanth , Agnihotri Sourav , Mehta Preeti , Salve Suneeta , Yoganand Raksha , Tarai Bansidhar , Tejashree A , Hemashettar Basavaraj , Balajee G , Gopal Lokeshwari , Gupta Sunita , Ghosh Anup , Rudramurthy Shivaprakash
Published: September 2022

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

 

Objective

Rhytidhysteron spp. are emerging dematiaceous fungi implicated in subcutaneous infections in immunocompromised as well as immunocompetent individuals. Diagnosis of Rhytidhysteron spp is delayed owing to non-sporulation and misidentification. The present study was conducted to characterize Rhytidhysteron spp. isolated from India by molecular techniques.

Method

We retrieved a total of eight isolates of Rhytidhysteron spp from our National Culture Collection for Pathogenic Fungi (NCCPF) that were received from various centers across India. Colony morphology on Sabouraud's dextrose agar (SDA) and potato dextrose agar (PDA) was recorded. Molecular identification was performed by sequencing internal transcribed spacer (ITS) and 28 s region of rDNA. Phylogenetic tree was constructed using the Neighbor-Joining method along with sequences of standard strains retrieved from the NCBI database.

Results

Rhytidhysteron spp was confirmed by sequencing from cutaneous specimens of eight cases. Six patients were immunocompetent while two were post-renal transplant recipients. Phenotypic features on SDA and PDA revealed growth of greyish-black mycelia on the obverse with black pigmentation on reverse after 7 days of incubation. On microscopic examination of lactophenol cotton blue mount, only dematiaceous septate hyphae without any spores were noted. On phylogenetic analysis of 28 s region, four of our isolates were closely clustered with Gloniopsis calami (Fig.1b), whereas two isolates with G. pneumonia and one each with G. percutanea, and Gloniopsis spp. (Fig.1b). On ITS phylogeny, the same four isolates closely clustered with G. calami (Fig.1a) as in 28 s region, one each isolate clustered with G. pneumonia, and G. percutanea while two isolates formed a separate clade with Rhytidhysteron rufulum (Fig.1a).

Conclusion

This fungus is often difficult to identify due to lack of sporulation making morphological identification challenging. Therefore, molecular sequencing is a must for its identification. However, identification using the sequence of ITS and 28 s rDNA does not clearly correlate. Hence to confirm the identity additional genes needs to be sequenced and analyzed.
P401 Identification and characterization of cryptic species of Aspergillus isolated from clinical samples
Authors: Janani R. Sruti , Singh Gagandeep , Xess Immaculata , Das Bimal Kumar
Published: September 2022

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

 

Background and Objective

Molds are emerging as a major cause of life-threatening infections in immunocompromised patients. There is an increasing recognition of the cryptic Aspergillus species, which are organisms that are morphologically indistinguishable yet can be differentiated by molecular methods. These organisms have been known to show a higher minimal inhibitory concentration for the majority of the antifungal agents in vitro. Therefore, correct identification of these cryptic species is very important to administer a proper antifungal agent.

In this study, we wish to identify and characterize the cryptic species of Aspergillus from all clinical samples.

Methods

Patient routine samples like broncho alveolar lavage, endo-tracheal aspirate, sputum, pus, tissue, and CSF from various wards, OPDs, and ICUs of All India Institute of Medical Sciences, New Delhi which were processed in the Mycology laboratory, department of Microbiology showing growth of Aspergillus species were included in this study. Identification of the isolates was done using phenotypic methods and by Matrix Assisted Laser Desorption Ionization-Time of Flight (MALDI-TOF). For the analysis of cryptic species, the isolates underwent PCR and then sequencing of the ß-tubulin gene was done. Antifungal susceptibility testing was done using micro broth dilution as per the CLSI method.

Results

Of the 92 isolates, using morphological methods, 53 were identified as A. fumigatus, 33 were identified as A. flavus, 3 as A. nidulans, 2 as A. terreus, and 1 as A. niger MALDI-TOF (Vitek MS database) misidentified 2 isolates of A. nidulans and 1 isolate of A, stellatus as A. fumigatus and 1 isolate of A. tubingensis as A. niger. The ß-tubulin sequence analysis for the identification of cryptic species revealed that 2 isolates (2.08%) were cryptic, one was A. stellatus morphologically identified as A. nidulans and another one was A. tubingensis morphologically identified as A. niger.

Conclusion

Currently the available data on cryptic Aspergillus species is very limited. Because of its varied susceptibility pattern, it is important to identify the Aspergillus isolates to its species level. There is also a need for expansion of the number of strains for each species in MALDI-TOF MS databases for convenient, faster, and correct identification.
P409 Study of epidemiology, risk factors and antifungal sensitivity pattern of fungal pneumonia in critically ill cirrhotics
Authors: Singh Paras , Kale Pratibha , Khillan Vikas , Sarin Shiv Kumar
Published: September 2022

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

 

Objectives

Liver cirrhosis causes immune dysregulation and increased susceptibility to fungal infections. We studied the epidemiology and risk factors, and compared the rapid diagnostic methods and biomarkers for fungal pneumonia in critically ill cirrhotics.

Methods

Single-center, prospective cohort study of 100 critically ill cirrhotics with fungal pneumonia between January to September 2021. Comparative analysis was done for culture, realtime PCR and biomarkers, bronchoalveolar lavage and serum Galactomannan, and serum procalcitonin measured on days 1, 3, and 7. The final outcome considered was mortality within 1 month after diagnosis or discharge.

Results

Aspergillus flavus was the most common species (70/100,70%). Risk factors were, neutropenia (P .03), steroids prior to ICU admission (P .02), prolonged hospitalizations ˃21 days (P .05), and culture positivity was 80%. The culture was not inferior to realtime PCR for the diagnosis of fungal pneumonia. BAL Galactomannan was an early prognostic marker with a median rise above ˃ 3.5 index value. The Median PCT level was higher from day 1in the fungal pneumonia non-survivor group (3.29 vs. 0.8 ng/ml) with higher 30-day mortality (72%). Higher PCT was associated with bacterial co-infection (48%), antibiotic (74%), antifungal therapy, and renal failure and mortality.

Conclusion

Fungal pneumonia complicates cirrhotics with neutropenia, prolonged hospitalization, and steroids as risk factors. Aspergillus flavus predominate in consensus with Asian epidemiology. Culture methods are reliable and a combination of molecular tests with BAL Galactomannan is useful for rapid diagnosis. Serum PCT is raised in patients with fungal pneumonia and is associated with higher mortality. In our study the baseline PCT at admission to ICU was higher in the non-survivor group, and levels on D3 and D7 were persistently higher. High serum procalcitonin level is an independent prognostic biomarker of mortality risk in fungal pneumonia.
P412 QuicGM Aspergillus Galactomannan Ag lateral flow assay as a practical aid to diagnose invasive Aspergillosis in high-risk hematology patients
Authors: Aerts Robina , Mercier Toine , Maertens Johan , Lagrou Katrien
Published: September 2022

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

 

Background

Early diagnosis of invasive aspergillosis (IA) is an important factor to improve survival, but it remains challenging. QuicGM Aspergillus Galactomannan Ag Lateral Flow Assay (LFA) is based on a fluorescence immunochromatography test using a monoclonal antibody to identify Aspergillus galactomannan antigen in serum and bronchoalveolar lavage fluid.

Methods

In this study, we analyzed the diagnostic performance of QuicGM in 120 serum samples of high-risk hematology patients [41 cases of proven (n = 4) and probable (n = 37) IA and 79 equally high-risk controls] and compared it to performance of the galactomannan (GM) [Platelia Aspergillus enzyme immunoassay (ELISA) (Bio-Rad, Marnes-la-Coquette, France)]. Probable cases defined by a positive Biorad GM in serum were also included in the cohort.

Results

Using the same cut-off of 0.5, QuicGM was positive in 22 cases versus GM in 21 cases. However, QuicGM was false positive in 11 controls with GM only in 1 control. These false positive samples remained positive upon repeated testing with the QuicGM. The test results of QuicGM and GM correlated well with a Spearman's correlation coefficient of 0.80 (P <.001). McNemar test showed no significant difference between both tests (P = 1) and Cohen's kappa coefficient was 0.65 (CI 0.49-0.81), suggesting a substantial strength of agreement.

QuicGM sensitivity (using 0.5 cutoff) was 54% versus 51% for GM. Specificity was 86% for QuicGM versus 99% for GM. Positive predictive value (PPV) was only 67% for QuicGM versus 95% for GM. Combining GM and QuicGM in serum increased sensitivity to 59%. Youden index of QuicGM in the cohort was 0.39. Using this cutoff, sensitivity was 66% and specificity was 80%. When using a higher cutoff of 0.6 for the QuicGM, specificity increased to 94% and PPV to 78%, at the cost of a decrease in sensitivity to 44%.

Discussion

Including probable cases defined by a positive Biorad GM in serum could have led to a bias in favor of the GM.

With the cutoff suggested by the company, there was a notable decrease in specificity and PPV for the QuicGM. The need to adjust this cut-off must be questioned. With the implementation of LFAs in the EORTC-MSGERG criteria, one usually strives for the same cutoff of 0.5, to assure a practical approach. However, we must remain thoughtful about slight differences in these tests. They all have their own IgG and IgM built. Caution is needed more in the setting of diagnosis than in screening.

More knowledge must be obtained regarding how different hospital laboratories have implemented the LFAs.

Conclusion

Diagnostic performances of QuicGM and Biorad GM correlate well.

Further research is necessary regarding the optimal cutoff index.
P435 Evaluation of remote triazole capillary blood testing to facilitate remote therapeutic drug monitoring (TDM): A validation study
Authors: Nwankwo Lisa , Donovan Jackie , Ni Melody , Loebinger Michael , Armstrong-James Darius , Shah Anand
Published: September 2022

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

 

Background

The advent of COVID-19 has meant that patients with chronic diseases needed to shield, however, investigations were needed to guide continual management of their disease. Remote monitoring options were evaluated to ensure the standard of care is not compromised.

Purpose and Hypothesis

The aim was to validate remote (return-posted) capillary triazole blood testing and evaluate the potential role of remote TDM in chronic antifungal therapy.

Materials and Methods

A single-center prospective cross-sectional study of remote finger prick capillary blood testing compared with gold standard venesection was performed. Remote finger prick capillary blood testing was validated compared to local standard venesection using comparative statistical analysis Comparative statistical analysis: Paired t-test, correlation and Bland-Altman were used to determine if there was agreement or association between the sampling methods.

Results

A total of 66 patients receiving triazole therapy were recruited and 57 pooled pairs of remote capillary and venous triazole concentrations and metabolites were prospectively analyzed, with the rest of the blood samples not being analyzed due to insufficient sample, hemolysis, or undetectable triazole level of <0.2 mg/l. There was a significant difference in the comparison of the two methods of sampling with paired t-test at P <.0001. Bland-Altman analysis yielded wide bias (−49.07%) and wide limits of agreement (−85.5% to −12.64%). On average capillary triazole, concentrations were 37% lower than venous concentrations (Fig. 1). There was however a very strong correlation between capillary and venous tests (Pearson's correlation coefficient r = 0.9219, P <.0001, Fig. 2).

Conclusions

Remote capillary triazole sampling does not appear interchangeable with venous sampling, but being strongly correlated and on average 2/3rd of the venous value, could be a predictor of venous triazole level, or be useful for intra-patient longitudinal monitoring. When incorporated into an outpatient clinical pathway it can improve shared decision-making and patient experience. Further research is required to determine appropriate target reference ranges if the new lower capillary levels can be used routinely, especially in the climate of COVID-19 where social distancing measures limit patient access to hospitals and clinics for routine investigations.
P441 Molecular identification, genotyping, and antifungal susceptibility of Trichosporon species isolated from clinical samples of patients at various parts of the Indian subcontinent
Authors: Parashar Abhila , Rastogi Vijaylatha , Prakash Hariprashad , Rudramurthy Shivaprakash M.
Published: September 2022

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

 

Objectives

(1) To study mycological characteristics of strains belonging to Trichosporon and its related genera obtained from clinical samples of patients from India. (2) Molecular identification by intergenic spacer (IGS) region 1 sequencing of the rDNA locus. (3) Genotyping of the major causative agent, T. asahii, and its in vitro drug susceptibility testing.

Materials and Methods

A total of 55 clinical isolates of Trichosporon species were collected from NCCPF (National culture collection of pathogenic fungi) PGIMER, Chandigarh along with different health institutions of India. These isolates were recovered from urine, blood, sputum, nail, tissue biopsy, pleural fluid, hair, BAL, and wound discharge over a period of 12 years (2006–2018). The isolates were molecularly characterized and genotyped using IGS-1 region sequencing. In vitro drug susceptibility testing of the isolates was performed against amphotericin-B, fluconazole, itraconazole, voriconazole, and posaconazole according to the CLSI M27-A3 guidelines (CLSI 2008).

Results

Predominant underlying risk factors identified were presence of an indwelling catheter, use of broad-spectrum antibiotics, and presence of comorbid conditions such as diabetes, hypertension, and anemia. A total of 47 (85%) of the 55 isolates were identified as T. asahii, 6 were T. inkin (11%), and 2 were Cutaneo Trichosporon dermatis (3.6%). Trichosporon asahii genotype III (22; 41%) was the most common type, followed by genotype IV (12; 22%), I (8; 15%), and VII (2; 4%). In addition to the 15 known T. asahii genotypes, one novel genotype was identified in this study. Indian T. asahii isolates showed high MIC ranges to amphotericin B (0.06-4 μg/l) and fluconazole (0.25-64 μg/l). Relatively low MIC ranges were found in the case of voriconazole (0.03-1 μg/l), Posaconazole (0.06-1 μg/l), and itraconazole (0.06-1 μg/l). Voriconazole appeared to be the most active drug in maximum T. asahii isolates. The MICs for all the drugs were comparatively lower in the case of non-T. asahii strains.

Conclusion

Trichosporon asahii remains the most common etiology of Trichosporonosis in India and presents a challenge for both diagnosis and treatment. With increasing drug resistance, therapeutic options are limited, and antifungal regimens with triazoles especially voriconazole appear to be the best. Accurate timely identification, removal of indwelling catheters/central venous lines, and voriconazole-based treatment along with control of underlying conditions were associated with favorable outcomes. Identification of the novel genotype has epidemiological implications and requires further work up.
P455 Identification of Cryptic species of Aspergillus using Beta-tubulin gene in a tertiary care center in South India
Authors: Karunai Almas Fathima Upaisal , Jyoti Kindo Anupma , Kumar S. Prasanna , Thirunarayan M. , Sree V. Lakshmi
Published: September 2022

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

 

Objectives

To identify cryptic species of the genus Aspergillus using Beta-tubulin gene by sequence typing.

Methods

Aspergillus grown from various clinical samples (Ear swab, Bronchial wash, Endotracheal Aspiration, Paranasal sinus, BAL, Sputum) was subcultured on Sabouraud's Dextrose Agar/Oatmeal Agar.

Tease mount/Slide culture was done to study the morphological features of the hyphae, size, shape, and arrangement of the conidia.

DNA Extraction using phenol-chloroform method: DNA was extracted and purified directly from 4-day-old culture and used as a template for polymerase chain reaction (PCR) amplification.

DNA Amplification: A fragment of the target gene were amplified using PCR.

The primer pairs used for Beta-tubulin gene was Bt2a (Forward primer) and Bt2b (Reverse primer).

Beta-tubulin.

DNA Purification: The PCR product was purified with multi-screen filter plates.

Gel electrophoresis: The amplified DNA product was subjected to agarose gel electrophoresis and specific band formation was to be observed for the species of Aspergillus.

DNA Sequencing: The purified product will be used as a template for sequencing. An applied biosystem 3730 sequencer will be used to obtain DNA sequences.

Phylogenetic Analysis: Parsimony analysis of individual and combined matrices will be conducted using PAUP version 4.0b10 software/free software at phylogeny.fr by the maximum parsimony (MP) and Neighbor-joining methods.

The results will be represented using a dendrogram.

Results

A total of 30 Aspergillus isolates were collected and identified using Phenotypic methods.

The DNA extraction and PCR amplification were done and sent for Sequencing. The results of Sequence typing are awaited.

Conclusion

Cryptic species are morphologically indistinguishable forms of Aspergillus and their identifications can be confirmed exclusively by using molecular techniques which have led to the description of previously unknown or rare species among different Aspergillus species complex. According to studies, the frequency of such species was found to be 10% in TRANSNET study U.S and 12% in FILPOP study Spain. Most of these species are less susceptible and certain species are multidrug-resistant (azoles and amphotericin B) and the emergence of antifungal-resistance among the Aspergillus species forms the importance in identifying Cryptic species.
P460 The Aspergillus Lateral Flow Assay for the diagnosis of invasive Aspergillosis
Authors: Liu Chonglong , Peng Jie , Zhang Yuan , Wang Ying , Wang He , Su Yan , Zhou Zeqi
Published: September 2022

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

 

Objectives

Invasive Aspergillosis (IA) is a life-threatening opportunistic mycosis that occurs in some people with a compromised immune system. The detection of Aspergillus galactomannan (GM) rapidly gained widespread acceptance as part of the diagnostic workup of a patient suspected of IA. Recently, a Novel QuicGM™ Aspergillus Galactomannan Ag Lateral Flow Assay (LFA) from Dynamiker Biotechnology (Tianjin) Co., Ltd. was released as a screening test of IA. It is a fluorescent immunochromatographic cassette test which using a monoclonal antibody against GM and Europium nanoparticles (Eu NP) (Fig. 1). It is a semi-quantitative test which was easy to operate, rapid with portable detection devices, and can be widely accepted by clinical and primary medical (Fig. 2). We conducted a comparative study between the Dynamiker QuicGMTM Aspergillus Galactomannan Ag (LFA) and the Bio-Rad Platelia™.

Methods

A total of 66 serum samples and 54 BALF samples were collected, 15 serum samples from the College of American Pathologists (CAP) certificated lab were also included in this study. The samples were tested in parallel using the Bio-Rad Platelia™ Aspergillus Ag (ELISA) and Dynamiker QuicGM™ Aspergillus Galactomannan Ag (LFA).

Results

The total coincidence rate of the two assays was 98.48% (Kappa value was 0.97 >0.75) and 90.74% (Kappa value was 0.81 >0.75) in 66 serum samples and 54 BALF samples respectively.

Conclusions

The performance of the Dynamiker QuicGM™Aspergillus Galactomannan Ag (LFA) and Bio-Rad Platelia™ Aspergillus Ag (ELISA) were highly consistent, and the performance of Dynamiker Aspergillus Galactomannan kit were all in line with the statement of the manufacturer. The results are accurate and reliable, and it could be used as an aid for the early rapid screening test of IA.
P479 Causative agents of Onychomycosis and associated factors: A 15 year study
Authors: Genç Gonca Erköse , Nuriyev Kamran , Küçükkaya Sertaç , Şatana Dilek , Uzun Meltem , Erturan Zayre
Published: September 2022

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

 

Objectives

Onychomycosis is a term used to describe nail infections that can cause discoloration, thickening, and separation from the nail bed. These manifestations can cause pain and discomfort and medical intervention may be required. The treatment of this disease is difficult and a time-taking process.

The aim of this study was to determine the frequency and distribution of fungal agents according to age and gender in patients with onychomycosis who applied to our laboratory over a period of 15 years.

Methods

A total of 1337 nail samples, which were sent to the Mycology Laboratory of Istanbul Medical Faculty Hospital, Department of Medical Microbiology between January 2007 and July 2022, were examined with conventional methods.

Calcofluor white and 15% KOH were used for microscopic examination and Sabouraud dextrose agar containing antibiotics with and without cycloheximide was used for culture. Fungi were identified by using conventional methods and/or API, VITEK, or MALDI-TOF MS.

Results

The distribution of 1337 nail samples according to patients’ body region, gender, and age is shown in Table 1. Of the total samples, 945 and 392 were toenails and fingernails respectively. Of the specimens, 1212 (90.6%) were taken from adults and 125 (9.4%) were taken from pediatric patients.

Fungal growth was detected in 139 (11.9%) specimens and one culture showed mixed growth. Yeasts were found to be the most frequent isolated fungi (60.7%) and dermatophytes were at the second place (25.71%). Among yeasts, non-albicans Candida species (76.4%) were found to be more frequent than Candida albicans (17,6%), Trichophyton species were found to be the most frequent dermatophytes (75%). Candida parapsilosis was the most prevalent among identified non-albicans Candida (21.2%). The distribution of fungal agents according to patients’ body region and age is shown in Table 2.

Among the patients with a positive culture, the most frequent underlying comorbidities were diabetes in 16 patients (11.5%) and HIV infection in 4 patients (2.87%). No significant difference was found when culture positivity rates were compared according to age and gender. However, there was a significantly higher rate of culture positivity in fingernails compared to toenails (P < .05).

Conclusion

Onychomycosis agents differ according to regions. In our analyses, we found non-albicans Candida species, especially C. parapsilosis as the most frequent agents of onychomycosis. Because these species are mostly resistant to antifungals this must be kept in mind during therapy.

As comorbidities were only found in a limited number of patients additional studies are needed to find responsible factors.
P493 Functional genomics in Candida glabrata, new tools to study stress, pathogenesis and drug resistance
Authors: Usher Jane
Published: September 2022

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

The processes of life are dynamic and changes on a molecular level enable organisms to grow but to adapt and survive in different environments, such as the ability to cause disease within a human host. My research focuses on the human fungal pathogen, Candida glabrata, which can cause illnesses in humans ranging from allergic reactions, infections such as thrush which affects ∼75% of women at least once, to serious diseases in patients that have impaired immune systems. These fungi are increasing in incidence and the reason for this increase is not understood. However, it is clear that the fungus can defend itself against high levels of stress and antifungal drugs used in treatment regimes. My hypothesis is that C. glabrata has evolved the capabilities to withstand a challenge from the combination of environmental and imposed drug stresses.

 

To look at C. glabrata, I will take advantage of my recent discovery of the sexual cycle in this fungus which offers novel methods to test hypotheses about evolution and pathogenesis. Pathogens of humans, such as C. glabrata, are successful because they adapt effectively to environmental stresses encountered within the host body. Upon recognition by host immune cells, C. glabrata is engulfed and exposed to a combination of stresses. In contrast to other pathogenic fungi, C. glabrata is highly resistant to stress allowing it to survive the host immune defenses. This suggests that resistance to both antifungal drugs and natural host-induced stresses are essential for the establishment and progression of infection. The molecular mechanisms underpinning antifungal resistance and the response to individual stresses have been investigated in isolation, however, little is known about how C. glabrata adapts to combinatorial stresses. The mechanistic explanation of stress adaptation will yield new insights into Candida infection.

 

Using my newly discovered sexual cycle in C. glabrata, I have generated a series of related strains of the same fungal pathogen that have increased resistance to combinatorial and drug stresses. I will sequence their genomes to identify the critical genes involved in stress resistance and characterize the mechanisms of C. glabrata stress responses. My preliminary data demonstrate that the C. glabrata response to in vitro combinatorial stress is similar to that observed upon phagocyte engulfment. At the level of gene expression, there is an up-regulation of genes encoding functions related to stress adaptation and nutrient recycling overlap. Understanding this regulatory network and the role that selected components (different genes) play in stress resistance, is essential to the development of future drug regimes.
P498 Pneumocystis Jirovecii pneumonia: Clinical profile and diagnostic modalities
Authors: Tarai P Bansidhar
Published: September 2022

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

 

Objectives

In developing countries like India, the diagnosis of Pneumocystis jirovecii infection is often made either by conventional staining or clinically. This study was planned to know the utility of polymerase chain reaction (PCR) in diagnosing Pneumocystis jirovecii pneumonia (PJP), to compare the PCR results with that of the staining technique, and also to correlate the results with the radiological condition of patients from a tertiary care center in India.

Method

A retrospective study included 3500 adult in-patients with symptoms of lower respiratory tract infection. Sputum and bronchoalveolar lavage samples were proceeded for both staining and PCR for multicopy mtLSU gene and singlecopy DHPS fas gene of P. jirovecii.

Results

The present study was carried out in the Department of Microbiology, Max, Hospital, Saket, New Delhi, India. Between the years 2019 and 2022, the patients with clinically suspected pneumocystis pneumonia who attended outpatient or admitted to the Departments of Medicine, Gastroenterology, and Neurology, etc. of our hospital were retrospectively analyzed. The details of these patients were analyzed as per a well-structured proforma that included the detailed clinical history and laboratory data from the hospital records. A total of 3500 patients were included in the present study. Of these 3500 patients, 1.42% (50/3500) patients of pneumocystis pneumonia were confirmed, based on positivity by at least any two of the above-mentioned techniques. The age of the patients ranged from 29 to 80 years (mean 32 years). Among these 50 patients, 34 (68%) were males and 16 (32%) were females and the male-to-female ratio was 2:1. A total of 10 patients had human immunodeficiency virus (HIV) infection, while the other 25 were HIV negative. The immune status of five patients was unknown. Fever and dyspnea (n = 44, 88%), non-productive cough and abnormal auscultation sound (n = 30, 60%), and also chills and weight loss (n = 10, 20%) were the documented clinical symptoms of PJP. Radiological diagnosis of Pneumocystis pneumonia was noticed in 34 cases. Foamy alveolar casts were present in 19 cases. Silver methenamine stain enhanced the rounded, helmet, or cleft forms of sporozoites. Inflammatory infiltrate was mainly polymorphonuclear. All the cases were confirmed as P. jiroveci by PCR amplification.

Conclusion

Clinicians often face the diagnostic dilemmas presented in the study. Individual modalities of the diagnosis are available, but all have drawbacks with varied sensitivity and specificity. Combining the available clinical, radiological, and microbiological modalities to reach early diagnosis can go a long way to avoid misdiagnosis and unnecessary delays in treatment.
S1.5c Proteomics in fungal keratitis research: a road map to personalized treatment
Authors: Prajna Lalitha
Published: September 2022

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

Research becomes very significant and meaningful when it addresses a significant public health problem of a region. Fungal keratitis, ulceration of the cornea due to fungal infection, is one such serious problem. This infection that results in monocular blindness affects primarily the agrarian population and is considered to be a silent epidemic in India.

 

The current treatment for fungal keratitis is the topical application of antifungal drugs such as natamycin and voriconazole. Nearly 40% of the patients do not respond to this treatment and require corneal transplantation. Treatment with antifungal drugs is not a holistic approach as it addresses only killing the fungus. The exaggerated inflammatory response at the site of infection may be a crucial factor in the disease outcome. And, this is not taken into account during the treatment due to the lack of knowledge of the host response during the fungal infection. This was the starting point of our research in fungal keratitis—to understand the corneal immune response to fungal infection.

 

Using mass-spectrometry-based proteomics studies on a tear from keratitis patients, we identified that in response to the fungal infection, the complement and coagulation pathways were activated along with neutrophil-mediated defense responses, notably the neutrophil extracellular traps. These pathways and their cross-talk with each other were primarily responsible for the exaggerated immune response at the site of infection. We selected five tear proteins that were significantly altered and validated them to serve as indicators of the inflammatory status of the ulcer in keratitis patients. Further, we developed a predictive logistic regression model that incorporates tear biomarker levels and ulcer characteristics to identify the subset of patients who are unlikely to respond to the antifungal treatment. We are currently exploring the possibility of using tear-derived EVs or their cargo as adjuvant therapy to modulate the inflammatory response in these non-responder patients.

 

Through our efforts using proteomics approaches, we now have five tear proteins as indicators of the inflammatory status in keratitis patients. These proteins along with the clinical features can identify the subset of patients who are unlikely to respond to antifungal treatment. Additionally, we showed that keratitis patient tear-derived extracellular vesicles are enriched with proteasomes. As proteasomes have an established role in immune modulation, EVs with proteasomes are thus promising candidates for adjuvant therapy, which we are currently exploring. Thus, our journey of over a decade of research on fungal keratitis started with the basic research to understand the host response that in turn provided the leads for translational research, which is now advancing towards personalized treatment for these patients.
S2.4d Combination antifungal effects of eugenol with voriconazole against Candida tropicalis and Candida krusei strains isolated from the genital tract of mares
Authors: Sharifzadeh Aghil , Shokri Hojjatollah , Nikaein Donya
Published: September 2022

S2.4 Veterinary mycology research, September 21, 2022, 3:00 PM - 4:30 PM

 

Objectives

The objective of the current study was to investigate the possibility that eugenol synergizes the antifungal effects of voriconazole on genital isolates of Candida krusei and Candida tropicalis from mares.

Methods

The antifungal activity of eugenol and voriconazole was evaluated using the broth microdilution assay (CLSI- M27-A3). The synergism of eugenol and voriconazole against genital Candida isolates was evaluated by the microdilution checkerboard method.

Results

Minimum inhibitory concentration (MIC) values for eugenol and voriconazole ranged from 400 to 800 μg/ml and 1 to 8 μg/ml, respectively, for C. tropicalis isolates, and from 200 to 400 μg/ml for eugenol and 2 to 16 μg/ml for voriconazole against C. krusei isolates. Eugenol decreased the arithmetic mean of MIC for voriconazole against C. tropicalis and C. krusei isolates from 2.66 to 0.46 μg/ml and 7.77 to 0.41 μg/ml respectively. The fractional inhibitory concentration index (FICI) values for the eugenol voriconazole combination ranged from 0.25 to 0.88 and 0.19 to 0.63 for C. tropicalis and C. krusei isolates respectively. A synergistic effect of eugenol in combination with voriconazole was observed for 83.3% of C. tropicalis and 77.7% of C. krusei isolates. The antagonistic activity was not seen in any of the isolates tested.

Conclusions

Eugenol showed fungistatic and fungicidal effects against genital Candida isolates and, in combination, synergized the antifungal effects of voriconazole. The eugenol-voriconazole combination can lay the foundation for a therapeutic approach against isolates in which azole resistance has increased over time.
S3.4d The role of NLRP3 inflammasome in host defense during Talaromyces marneffei infection
Authors: Sha Lu , Jinquan Wu , Liyan Xi , Zhou Xin
Published: September 2022

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

Talaromyces (Penicillium) marneffei (T. marneffei) is the only thermally dimorphic pathogen in Talaromyces. The pathogenesis of T. marneffei in mammals is not yet fully understood. Inhalation of T. marneffei conidia without normal clearance may result in conidia dissemination throughout the body and lead to disseminated infection. In TSM patients, study have shown that IL-1β and IL-18 levels increased and were consistently associated with the severity of sepsis and outcomes of post-treatment. That means poor outcome likely was associated with an overly strong immune response. Several studies have identified inflammasome activation as an essential immune response in host defense against fungal pathogens. Among them, NLRP3 inflammasome is the most widely characterized. However, the role of NLRP3 inflammasomes in T. marneffei-induced immunopathology remains to be elucidated. Therefore, in the present study, we aimed to address the role played by the NLRP3 inflammasome in the T. marneffei systemic infection in mice.

 

We established T. marneffei infected murine pulmonary model with two groups of mice, including the Nlrp3-/- mice and wild-type mice.

 

We found that infected mice displayed NLRP3 inflammasome activation and increased production of IL-1β upon pulmonary T. marneffei infection. Further, we demonstrated that T. marneffei conidia activated the NLRP3 inflammasome both in mice and human macrophages. And T. marneffei conidia induced IL-1β released by infected macrophages is NLRP3 inflammasome-dependent. In vivo study, we found that NLRP3 contributes to the development of lethality in the early stage of pulmonary T. marneffei infection. However, Nlrp3-/- mice showed a similar fungal load to the WT in the middle stage of infection and a significantly increased number of fungi recovered from the lung of the WT mice could be seen in the late stage of infection. Moreover, NLRP3 contributes to pathogenic inflammation in pulmonary T. marneffei infection and contributes to neutrophil recruitment and pulmonary injury.

 

So, in the present study, we demonstrated that the NLRP3 inflammasome is activated during T. marneffei infection. But NLRP3 inflammasome plays a dual role during pathogenic T. marneffei: an early inflammatory response inducing a protective environment, and a subsequent excessive damaging inflammatory response that contributes to pathogenesis and mortality. This study identifies for the first time that activation of the inflammasome in the later stages of TSM detrimentally contributes to pathogenesis and suggests that targeting the inflammasome may be a therapeutic option to treat pathogenic T. marneffei infections.