Fungal Culture Research Articles

Utility of Mucorales PCR to diagnose Rhizomucor infections in neutropenic patients.

Mucormycoses are life-threatening infections related to fungi from the Mucorales order. Based on fungal culture, the most frequently involved genera are Rhizopus spp., Mucor spp. or Lichtheimia spp. However, since the introduction of Mucorales PCR, many diagnoses are made without positive fungal culture, biasing mucormycosis epidemiology. We conducted a single-center retrospective observational study on invasive mucormycosis cases diagnosed between April 2020 and December 2022. Cases were classified according to EORTC/MSGERC definitions, adding a "PCR-only" category for patients with a positive Mucorales PCR as the only mycological evidence. Genus/species identification was obtained by sequencing the Mucorales 18S rDNA directly on Mucorales PCR-positive samples. We identified 35 cases of mucormycosis, including 6 proven, 7 probable and 22 "PCR-only". Genus/species identification was achievable in 34 cases, and surprisingly revealed the genus Rhizomucor as the main aetiological agent (n=14, 41.2%). Interestingly, all the Rhizomucor infections, except one, were classified as "PCR-only", while fungal culture was positive in 11/20 (55%) for other Mucorales genera (p < 0.001). Moreover, in comparison with other genera, the genus Rhizomucor was significantly more associated with neutropenia (11/14 [78.6%] vs 2/20 [10%], p < 0.0001) and pulmonary localizations (11/14 [78.6%] vs 6/20 [30%], p = 0.01). Our study reveals the changing epidemiology of mucormycosis in our center with the use of Mucorales PCR and underlines the importance of the genus Rhizomucor especially in neutropenic patients. This highlights the benefits using Mucorales PCR in clinical practice for mucormycosis diagnosis in high-risk patients, and the need to include it in diagnostic criteria.

First Report of Lasiodiplodia theobromae Causing Wilt and Fruit Rot of Pepper in Hainan Province, China.

Ornamental pepper (Capsicum annuum L.) is an economically important plant with extensive genetic diversity (Zhang et al. 2020). In September 2022, symptoms of wilt and fruit rot were identified in approximately 0.02 hectares of an ornamental pepper plantation in Haikou, Hainan Province, China (110°32' E, 20°06' N). Disease severity reached 85%, with an incidence rate of 90%. Symptoms started as black foliar spots that expanded into large lesions, spreading to fruits and stems, causing wilting (Fig. S1). Samples from symptomatic leaves, fruits, and stems of 27 plants were surface-sterilized with 75% ethanol for 30 s, rinsed five times with sterile water, air-dried, plated on potato dextrose agar (PDA), and incubated at 28°C for 5 to 6 days. To obtain pure fungal cultures, initial isolates were subcultured onto fresh PDA. Among the 27 fungal cultures, 11 isolates from leaves and fruits consistently formed gray to olivaceous colonies. Isolate LJY224 initially produced grayish-white, fluffy mycelia with radiating aerial hyphae. Over time, the colony became grayish-black and produced black, nearly spherical pycnidia. The conidia were oval, initially transparent, and single-celled. Mature conidia were dark brown, septate, with longitudinal striations, averaging 11.55 ± 0.75 µm in width and 24.93 ± 1.50 µm in length (n=35) (Fig. S2), indicating Lasiodiplodia spp. Genomic DNA was extracted using a fungal DNA extraction kit (OMEGA BIO-TEK, GZ Feiyang Biotech Co., Ltd., Guangzhou, China). Molecular identification involved sequencing the rRNA internal transcribed spacer (ITS) region and genes encoding β-tubulin (TUB) and translation elongation factor 1-α (TEF1) with primers ITS1/ITS4, Bt2a/Bt2b, and EF1-983F/EF1-2218R, respectively (White et al. 1990; Rosado et al. 2016; Rehner and Buckley 2005). BLASTn searches with the obtained ITS, TUB, and TEF1 sequences (GenBank accessions OQ612711, OR039814, OR039813) revealed 98% to 100% identity with Lasiodiplodia theobromae reference sequences from the NCBI database (OR018404, KR260830, MN461169), matching 541/548, 446/446, and 947/955 base pairs, respectively. A phylogenetic tree was constructed using concatenated multi-locus sequence analysis (MLSA) of ITS, TUB, and TEF1 (Fig. S3). To fulfill Koch's postulates, healthy 3-month-old ornamental pepper plants were inoculated by spraying the entire plant with a conidial suspension (50 ml, 107 conidia/L) of LJY224, using sterile water as a negative control. Each treatment included three replicates. Plants were maintained at 25°C, 75% humidity, under a 12-h light/dark cycle, and monitored daily. After 14 days, inoculated plants developed black foliar spots that expanded into large lesions, spreading to fruits and stems, and causing wilting, consistent with initial symptoms, while control plants remained healthy (Fig. S4). The re-isolated pathogen showed identical morphology to the original strain. In contrast, no fungi were isolated or recovered from the plants inoculated with water. Results from disease symptoms, colony and spore morphology, pathogenicity tests, and multi-locus DNA sequence analysis suggest that L. theobromae was the pathogen responsible for the disease symptoms on ornamental peppers. L. theobromae is closely related to Botryosphaeria dothidea, both of which cause fruit rot in peppers (Rui et al. 2023). To our knowledge, this is the first report of L. theobromae causing leaf wilt and fruit rot in ornamental pepper in Hainan Province, China, offering insights to mitigate crop losses.

“Curvularia Lunata-Induced Onychomycosis in an Unlikely Host - A Case report”

Introduction: Onychomycosis, a fungal infection of the nail apparatus, presents with various manifestations, including subungual hyperkeratosis, onycholysis and nail plate destruction. Established risk factors include trauma, advanced age, and comorbidities such as diabetes and immunosuppression. Trichophyton rubrum, classified as a dermatophyte, is the primary causative agent, with dermatophytes contributing to 60-70% of cases. Nondermatophyte molds (NDM) account for 30-40% of onychomycosis with Candida species being the most commonly identified, while Curvularia species are rarely encountered. Curvularia lunata, a saprobic dematiaceous mold, is an infrequent human pathogen primarily residing in soil. While human diseases caused by Curvularia are rare, they encompass a spectrum from skin infections to endocarditis. Case presentation: This case report details a 53-year-old male engineer presenting with left big toenail discoloration, revealing a Curvularia lunata-induced onychomycosis through positive KOH preparation and culture. Given the patient's concurrent use of ticagrelor, oral terbinafine hydrochloride was chosen as a therapeutic option, taking into account potential drug-drug interactions. This approach resulted in complete resolution. The discussion emphasizes distinctions in disease severity among immunocompromised and immunocompetent patients, the potential role of trauma in Curvularia infections, and the importance of comprehensive diagnostic examinations. Conclusion: This case report provides insight into the challenges of accurate diagnosis, emphasizing the need for routine fungal cultures. Additionally, it discusses the complexities of treatment options, considering factors such as drug interactions and patient-specific conditions, and highlighting the importance of tailored management plans. This case is reported because of the rarity of onychomycosis caused by Curvularia in immunocompetent individuals.

First Report of Leaf Tip Necrosis of Camellia oleifera Caused by Nigrospora aurantiaca in China.

In recent years, Camellia oleifera C.Abel. has become one of the most important woody oil trees cultivated at a large scale in Guizhou Province, China. However, the leaf diseases limited the development of C. oleifera industry. In October 2023, typical symptoms of leaf tip necrosis were observed on leaves of C. oleifera (Changlin series) at a plantation in Dongfeng Forest Farm (109°10'28'E, 26°18'41'N), Liping County, with disease incidence of 30-40% (n=220) in the observed fields (30 m × 30 m). The brown lesion started from the leaf tips, gradually extended inward then turned gray-white, curled, withered and fell off, and the boundary between the diseased and healthy parts was obvious. Symptomatic leaves (n=20) were collected and surface disinfected with 2% NaClO for 30 s, rinsed three times in sterile distilled water, air-dried and placed on potato dextrose agar (PDA) medium and incubated at 25°C for 7 d. The fungal colonies of 6 isolates were initially white, with red pigment, and later gradually turned gray, eventually turning black. Conidia were globose to sub-globose, black, smooth, single-celled, discrete and solitary, measuring 5.1 µm±1.28×4.7 µm±1.24 (n=50). The morphological features of the isolates are similar to the description of Nigrospora (Wang et al. 2017). For molecular identification, DNA of the representative isolate LP1021 was extracted using a fungal genomic DNA extraction kit (Sangon, Shanghai, China). The rDNA internal transcribed spacer (ITS), translation elongation factor 1-α (TEF-1α) and β-tubulin (TUB2) were amplified with the primers ITS1/ITS4 (White et al.1990), EF1-728F/EF1-986R (Carbone and Kohn, 1999) and Bt2a/Bt2b (Glass and Donaldson, 1995). The sequences of ITS, TEF-1α and TUB2 were deposited in GenBank as accession numbers PP499217, PP529954, and PP529955, respectively. BLAST revealed that ITS, TEF-1a and TUB2 sequences showed 100%, 98.80% and 100% similarity to those of N. aurantiaca (NR153477.1, KY019295.1 and MK388225.1), respectively. The phylogenetic tree constructed with the software MEGA X using the Neighbor-Joining algorithm (Felsenstein, 1985) indicated the isolate LP1021 separated from N. chinensis previously reported from C. oleifera in China. Based on morphological and molecular characteristics, the fungal pathogen was identified as N. aurantiaca. A representative isolate LP1021 was deposited in the Forest Protection Laboratory, Guizhou University. To further confirm the pathogenicity, a conidial suspension (1×104 spores/mL) of the isolate LP1021 was sprayed on six pots of two-year-old healthy C. oleifera seedlings in vivo. The same amounts of seedlings were sprayed with distilled water as control. All plants were placed at a constant room temperature of 25 ± 2°C. After 15 d, the inoculated leaves showed typical symptoms, similar to those observed in the field, while the control remained healthy. The re-isolated fungal culture was identical to that originally obtained using the methods mentioned above, which fulfilled Koch's postulates. To the best of our knowledge, this is the first report of N. aurantiaca causing leaf tip necrosis of C. oleifera in China, which provides an important theoretical basis for the pathogen identification and scientific control strategies of leaf tip necrosis in C. oleifera.

Wood's Lamp for Early Detection of Microsporum canis Tinea Capitis in Children

BackgroundPediatric alopecia is increasingly common, influenced by conditions like alopecia areata and infections such as tinea capitis, primarily caused by Microsporum canis. Diagnosing tinea capitis in primary care is challenging due to limited access to advanced tools, leading to treatment delays. ObjectivesThis study evaluates the effectiveness of Wood's lamp for early diagnosis of tinea capitis and its role in distinguishing it from other pediatric alopecia types. MethodsFifty pediatric patients under 12 with alopecia were enrolled from March 2023 to March 2024. Data collected included contact history, CFW microscopy, fungal cultures, Wood's lamp examinations, treatments, and follow-ups. ResultsWood's lamp identified 22 cases linked to animal contact, confirmed as Microsporum canis. Overall, 30 tinea capitis, 15 alopecia areata, and 5 trichotillomania cases were diagnosed. Post-treatment, fluorescence decreased in tinea capitis patients, who also showed hair regrowth, with no recurrence during follow-up. ConclusionsWood's lamp is a valuable, cost-effective diagnostic tool for tinea capitis, especially when combined with animal contact history, supporting its use in primary healthcare.

Assessment of pathogenic yeast species in the oral cavity of dogs: Antifungal susceptibility and implications for human health.

Companions' animals can present a risk for the transmission of opportunistic diseases to their owners, including those caused by yeasts of the Candida genus residing in their oral microbiota. This study aimed to isolate and identify yeasts from the oral cavity of dogs and assess their susceptibility to antifungals. Yeast species were identified using automated methods MALDI-TOF-MS and VITEK 2 from 50 dogs (aged 2-4years, various breeds). Among the evaluated animals Candida albicans (37.5%) and Candida parapsilosis (25%) were predominant, followed by other species including C. haemuloni, C. tropicalis, C. krusei, and Trichosporon asahii (12.5% each). In this study, antifungal susceptibility testing revealed a general susceptibility with limited resistance. Although most antifungals exhibited good efficacy, some Candida strains demonstrated resistance to fluconazole (FLC), caspofungin (CAS), and amphotericin B (AMPB). These findings highlight the importance of conducting fungal cultures and antifungigram testing for the effective management of fungal infections in humans and animals. Therefore, vigilant monitoring and control measures are essential to reduce the transmission of opportunistic diseases from pets to humans.

P-80. The microbiology of periprosthetic joint infections as revealed by sonicate cultures in Korea: Routine use of fungal and mycobacterial cultures is necessary?

Abstract Background Although sonication is a valuable diagnostic tool for periprosthetic joint infections (PJI), it is not commonly utilized. We analyzed sonicate and intraoperative tissue culture results obtained from three hospitals to define the microbial etiology of PJIs in Korea. Furthermore, we investigated necessity of conducting regular fungal and mycobacterial cultures. Methods We retrospectively analyzed data for patients with suspected orthopedic-related infections between 2017 and 2022, who had undergone prostheses removal surgery. We included 193 patients with suspected PJIs, and bacterial (n = 193), fungal (n = 193), and mycobacterial (n = 186) cultures were conducted on both sonicate and intraoperative tissue samples. The diagnosis of PJI was based on the European Bone and Joint Infection Society (EBJIS) criteria. Results Out of 193 patients, 121 (62.7%) had positive sonicate cultures, while 112 (58.0%) had positive periprosthetic tissue cultures. According to EBJIS criteria, a total of 181 patients were diagnosed with PJI, and 141 patients received microbiological confirmation through sonicate fluid culture or tissue culture. Of the 181 patients, 28 were classified with acute PJI (within 3 months of implantation) and 153 with chronic PJI. Among 141 patients, staphylococci were the most common organisms, accounting for 51.8% of cases, followed by Gram-negative organisms (15.6%), fungus (8.5%), and mycobacteria (3.5%). Nearly 91.7% of fungal isolates were Candida species, which also grew in bacterial cultures. In total, 11 cases cultured positive only in tissue culture, whereas 20 cases cultured positive only in sonicate culture. The antibiotic treatment plans were adjusted according to culture results. Conclusion Utilizing sonicate culture has greatly assisted in identifying pathogens responsible for chronic indolent PJIs, allowing suitable antimicrobial treatment. Based on few cases involving non-Candida and mycobacterial infections, it appears that routine fungal and mycobacterial cultures may not be necessary. Disclosures All Authors: No reported disclosures

P-2131. Identification of Non-fumigatus Aspergillus Species in Clinical Samples from Southern California

Abstract Background Aspergillus is the causative agent of aspergillosis, with presentations ranging from mild skin infections to disseminated disease affecting the central nervous system. Fungal culture from the infected site is the current recommendation to confirm presence of Aspergillus and to speciate based on morphology. A. fumigatus has historically been identified as the source of most serious infections, but there is emerging concern for invasive disease caused by non-fumigatus­ species, especially those that may carry intrinsic resistance to recommended antifungals. In this descriptive analysis, we reviewed trends in Aspergillus species cultured from a large clinical cohort.Figure 1.Percentage of Positive Aspergillus cultures by specimen origin type over time 2012-2023. Methods We identified all positive fungal cultures for at least one Aspergillus species across Kaiser Permanente Southern California hospitals and clinics from January 2012–June 2023. Episodes were defined as a combination of any Aspergillus culture results conducted within 30 days of the first positive Aspergillus culture. Only the first episode for each patient was included in analyses if multiple episodes were identified.Figure 2.Number of positive Aspergillus cultures by species identified and specimen type. Results 10,777 patients were identified at KPSC with at least one positive Aspergillus culture between 2012 and 2023. Isolates were most frequently identified from samples associated with the respiratory tract and bronchoalveolar lavage fluid. However, the proportion of cutaneous cultures has increased over time and was nearly equal to the number of positive respiratory cultures in 2023 (Figure 1). Aspergillus niger (n=3,772) was the most frequently cultured Aspergillus species overall, with 44.6% of isolates originating from skin, nail, and scalp samples (Figure 2). Aspergillus fumigatus was identified in 40.3% of respiratory sample cultures, followed by Aspergillus niger at 25.7%. Finally, non-fumigatus specimens exhibited seasonal trends in respiratory samples compared to A. fumigatus, peaking in early autumn each year (Figure 3).Figure 3.Number of positive Aspergillus cultures in respiratory samples by reported species by month. Conclusion Non-fumigatus species were identified more frequently than A. fumigatus in culture of respiratory and non-respiratory clinical samples. Novel observations of seasonal trends and increasing predominance of non-fumigatus species may indicate a shift in clinical expectations and treatment options during management of aspergillosis. Disclosures Emily Rayens, PhD, MPH, GSK: Grant/Research Support|Moderna: Grant/Research Support Sara Y. Tartof, PhD MPH, GSK: Grant/Research Support|Pfizer Inc: Grant/Research Support

Sjogren's Syndrome Complicated by Naso-Orbito-Cerebral Mucormycosis: A Case Report.

Background: Sjogren's syndrome (SS) is a chronic inflammatory autoimmune disease characterized by exocrine gland dysfunction. Mucormycosis is a rare yet life-threatening opportunistic fungal infection caused by Mucor species, with a high mortality rate. In patients undergoing long-term immunosuppressive therapy or corticosteroid use, especially when compounded by conditions such as diabetes or hyperlipidemia, Mucor can become pathogenic. Medical Record Description: We retrospectively analyzed the clinical data of a patient with Sjogren's syndrome complicated by rhinocerebral mucormycosis, focusing on its clinical presentation and progression. The patient was admitted to the hospital 20 days prior with symptoms of a "nasal infection" caused by a furuncle on the nose. The condition progressively worsened, resulting in nasofacial skin ulceration and necrosis, with black eschar formation around the affected area. Upon admission, comprehensive evaluations, including rapid histopathological and fungal tests, confirmed the diagnosis of mucormycosis. Despite surgical intervention under general anesthesia and antifungal therapy, the clinical outcome was ultimately fatal. Conclusion: Mucor infections commonly occur in patients with immunodeficiency and severe underlying diseases. The clinical manifestations are diverse, often presenting with skin tissue necrosis and the formation of black eschar. Fungal culture and histopathological examination remain the gold standards for diagnosing mucormycosis, while imaging studies are crucial for evaluating the extent of fungal dissemination and bone involvement. Given the rapid progression of the disease, early diagnosis and prompt treatment are critical for improving survival rates.

P-353. Aspergillus Burdens in Outdoor and Indoor Air at Hospitals Housing Patients at High Risk for Invasive Fungal Infections

Abstract Background Airborne dissemination of conidia is implicated in transmission of pathogenic fungi. There is lack of knowledge on airborne fungal burdens in hospital environments. Methods Air at hospitals A and B (1.5 miles apart) was sampled monthly (SASS 3100; 36000 L/2 hrs) from May ‘23-April ‘24 including outdoor, lobby, nurse stations (NS) and patient (pt) rooms on units caring for patients with immunocompromising conditions [Table 1]. New building construction was ongoing next to hospital A. Filter extracted DNA genome equivalent (GE) was quantitated by pan-Aspergillus (ASP) qPCR and standard curve; sonicated and non-sonicated filters were cultured. Descriptive statistics, Mann-Whitney U and Wilcoxon tests were used.Figure 1.Comparisons of Aspergillus burdens at 2 hospitals1A. Comparison of log ASP GE of outdoor air (outside of the entrance to the hospital lobby) over time at hospitals A and B (standard error shown in grey shade). 1B. Log ASP GE of air in the lobby of hospitals A and B. Note that ASP burden was higher in the lobby of hospital A, which is much more heavily trafficked than the lobby of hospital B. 1C. Log ASP GE of air at the nurses’ stations of 7 units in hospitals A and B. There is no significant difference between burdens at the 2 hospitals (p=0.74). 1D. Log ASP GE for rooms at hospitals A and B without HEPA filtration and positive pressure. Curves were plotted via locally estimated scatterplot smoothing (LOESS) with standard error shown in grey. Results Mean log-GEs were greatest for outdoor samples, followed by lobby, NS and pt rooms. Protected (HEPA-filtered and positive pressure) pt room GEs were significantly lower than unprotected rooms. There were no significant location-specific differences between hospitals A and B, except the lobbies [Fig 1]. GE from HEPA filtered rooms in hospital B unit 0 was taken as a comparative “safety threshold”, since filtering protects against infections. Extra GE burden in unprotected rooms was ∼0.5-1.2 log GE. A construction accident that exposed a unit in hospital A directly to outdoor air caused a ≥ 1-log GE spike that persisted from 2-4 weeks [Fig 2]. Seasonality data indicate outdoor peak and trough GEs in fall and spring, respectively. GEs were validated by + ASP and A. fumigatus air cultures [Fig 3].Figure 2.Trends of Aspergillus burden in units at hospital A Trends of log ASP GE over time for the patients’ rooms at hospital A ( A-1, A-2, A-3 units). Three new buildings were under construction within a block of hospital A. A-1 is the unit closest to the construction. Note the higher ASP burden in A-1 rooms compared with other units. Breach of the wall separating the outside environment and the patients’ rooms in A-1 occurred in late Nov (arrow). Note 1) the higher ASP GE in unit A-1 than other units even before the accident, and 2) the further increase in burden around the accident, which gradually came down to baseline after 2-3 weeks. Curves were plotted via LOESS with standard error shown in grey. Conclusion Airborne fungal exposure within hospitals is significantly lower than immediately outside the hospital. Fungal burdens within protected pt rooms can be used to estimate extra burdens encountered by immunosuppressed pts in unprotected rooms. Elevated fungal burdens stemming from contamination of a unit can persist for weeks following remediation. We are continuing studies of seasonality and correlations between airborne burdens, environmental and clinical fungal cultures, and fungal infections.Figure 3.Comparison of Aspergillus burden measured by qPCR and culture, stratified by locations3A. Mean log ASP GE for each location sampled. The reference line is the difference in mean log ASP GE when comparing that location to the B-0 rooms (HEPA-filtered, positive pressure). Note that the differences in log ASP GE between B-0 and other unfiltered units range from 0.52 to 1.2-log ASP GE. Upon entering the lobby, the log ASP GE was decreased by a median of ∼1 log, which was further decreased in patients’ rooms (3.16 to 4.42). 3B. Percent of positive cultures from each sampled location. AF designates cultures positive for Aspergillus fumigatus while ASP designates pan-Aspergillus¬ cultures. There was ∼30% decrease in positive cultures from outdoors to indoors, and an additional &amp;gt;10% decrease when moving to units. Culture positivity in unprotected NS and rooms was higher than protected NS and rooms by ∼10%. 3C. There is no significant difference between the means of GEs between the outdoors of the 2 hospitals (p=0.16). ASP burden was higher in the lobby of hospital A than B (p&amp;lt;0.001). There was no significant difference between means of the log ASP GE between the unfiltered rooms at hospital A and the unfiltered rooms at hospital B (p=0.67). Disclosures Alexander Sundermann, DrPH, CIC, FAPIC, OpGen: Honoraria Graham M. Snyder, MD, SM, Infectious Diseases Connect: Advisor/Consultant Cornelius J. Clancy, MD, Cidara: Grant/Research Support|Gilead: Honoraria|Merck: Grant/Research Support|Scynexis: Advisor/Consultant|Shionogi: Advisor/Consultant|Venatorx: Advisor/Consultant

P-1023. Prevalence and distribution of fungal microorganisms among cutaneous specimens submitted to community microbiology laboratories

Abstract Background Not much is known about the local prevalence of cutaneous fungal infections in Canada because these data are not readily available. Knowing the distribution of microorganisms in local settings may help developing guidance for empiric therapy, especially for immunocompromised patients whose cutaneous fungal infections could turn into systemic infections. The current retrospective study aimed to establish the prevalence and distribution of fungal microorganisms among patients who had specimens submitted to community microbiology laboratory for cutaneous fungal culture workups. Methods LifeLabs regional microbiology laboratories, connected with 129 collection centers in urban and rural communities in British Columbia, provided reports of isolated fungus from hair, nail, and skin scraping specimens from 1 January 2022 to 31 December 2022. Culture followed by microscopy and/or MALDI-TOF mass spectrometry was routinely used to isolate and identity microorganisms, but nucleic-acid testing was added to help identification if needed. Results Among the 20652 cutaneous specimens submitted in a year, 3665 of them were reported to have microorganism growth. The most reported microorganism was Trichophyton species (43.8%), followed by Penicillium species (5.8%), Candida albicans (4.2%), Aspergillus species (3.6%), and Fusarium species (2.8%). Of note, other than Trichophyton species, the other two main dermatophytes Microsporum species and Epidermophyton species accounted for only 0.2% and 0.1% of the positive specimens, respectively. Pseudomonas aeruginosa was incidentally identified in 2.9% of the positive specimens in the fungal culture workups. Conclusion Trichophyton species was the most reported microorganism among the hair, nail, and skin scrapping specimens submitted for cutaneous fungal culture workups from communities in British Columbia. Pseudomonas infections, which could present as chloronychia, folliculitis, ecthyma gangrenosum, could be considered in the differential diagnoses of cutaneous fungal infections. The current data will help clinicians to determine the empiric therapy needed based on each patient’s clinical situation. Disclosures All Authors: No reported disclosures

P-2136. Bronchoalveolar Lavage Mucorales PCR Testing in Immunocompromised Patients with and without Mold Infections

Abstract Background Early detection of mucormycosis is crucial for successfull treatment. Aim of the study was to evaluate the added clinical value of MucorGenius® PCR testing for detection of mucormycosis in bronchoalveolar lavage fluid (BALF) samples from patients at risk for invasive mold infections (IMI). Methods A total of 670 BALF samples collected between 2013-2023 from 638 patients were routinely tested for fungal culture and galactomannan before storage at -80°C. IMI was classified according to the FUNDICU 2024 criteria for intensive care unit (ICU) patients (n=277), or the EORTC/MSG 2020 criteria for patients outside the ICU (n=393) . BALF samples were retrospectively tested with the quantitative real-time MucorGenius® PCR assay between December 2023 and February 2024, and results compared to routine diagnostics and clinical classification Results Out of 670 BALF samples, 11 (1.6%) tested positive with the MucorGenius® PCR (mean ct value 32.95), 653 (97.5%) tested negative, while PCR was inhibited in 6 samples (0.9%). The 11 samples that tested positive included both samples with Mucorales growth in BALF culture. 3/11 samples showed growth of Aspergillus spp. (versus 29/653 samples that tested MucorGenius® negative), with BALF GM testing positive ( &amp;gt;0.5 ODI) in 5/5 samples tested, while serum GM was positive in 2/7 samples and Aspergillus PCR positive in 1/1 sample. Overall 8/11 samples had mycological evidence for IMI from routine diagnostics. Of 11 patients with positive MucorGenius® PCR, 6 (55%) met criteria of probable IMI (4 aspergillosis, 2 mucormycosis), while one met criteria of possible IMI. Of those with negative MucorGenius® PCR, 21/653 samples (3.2%) were classified as probable/proven aspergillosis. Therefore 4/25 (16%) of cases classified as probable/proven aspergillosis showed evidence of Mucorales co-infection when MucorGenius® PCR was applied. Conclusion Our study outlines the potential value of Mucorales PCR testing in BALF in patients at risk for IMI. The test was highly specific, detected both cases of probable mucormycosis, indicated Mucorales as causative pathogen in a patient with clinical and radiological evidence for IMI but negative routine test results, and detected mucormycosis co-infection in 16% of cases classified as probable/proven aspergillosis. Disclosures Martin Hoenigl, MD, Aicuris: Advisor/Consultant|Astra Zeneca: Honoraria|Gilead: Grant/Research Support|Gilead: Honoraria|IMMY: Grant/Research Support|Melinta: Grant/Research Support|Melinta: Honoraria|MSD: Grant/Research Support|Mundipharma: Grant/Research Support|Mundipharma: Honoraria|Pfizer: Grant/Research Support|Pulmocide: Advisor/Consultant|Pulmocide: Grant/Research Support|Scynexis: Advisor/Consultant|Scynexis: Grant/Research Support|Shionogi: Honoraria Juergen Prattes, MD, PhD, AbbVie Inc.: Stocks/Bonds (Private Company)|Gilead: Honoraria|MSD: Grant/Research Support|Novo Nordisk: Stocks/Bonds (Private Company)|Pfizer: Grant/Research Support|Pfizer: Honoraria

P-1038. Clinical Analysis of Nosocomial Candidemia: Species Profile, Resistance Patterns and Mortality Determinants

Abstract Background Candidemia has become one of the commonest nosocomial blood stream infection (BSI) secondary to infected central venous catheters or surgical site infections. Prompt diagnosis followed by early treatment is needed to reduce the high mortality associated with it. Hence this study was conducted to analyse the epidemiology, clinical characteristics, species distribution, resistance profile, outcome and factors related to outcome in nosocomial candidemia. Methods It was a retrospective observational analysis done for the period of 1 year in a tertiary health care centre of western Rajasthan from 1st July 2022 to 30th June 2023. All adult patients admitted in multispeciality intensive care unit (ICU) for more than 48 hours and developing candidemia were included and those patients with a single blood fungal culture positive from the sample obtained from indwelling central venous catheter were excluded as colonisers. Data was collected by regular AMS rounds and was analysed using SPSS version 20. Results Out of total 76 cases of candidemia,61 were included in the study. As shown in figure 1, the most common species isolated was Candida tropicalis followed by Candida parapsilosis.both were associated with high mortality. Clinicoepidemiological characters are described in table1. Overall, all cause 28th day mortality was 63.9 %. Highest resistance was found for the azoles. As depicted in table 2, on univariate and multivariate analysis infection with Candida tropicalis, use of inotropes, ventilator use, and high SOFA scores were the independent factors affecting mortality in candidemia. Conclusion Candidemia is a nosocomial infection with very high mortality which needs to be diagnosed early. Candida tropicalis being the most frequent species isolated in our ICU setup with a significantly high mortality. Disclosures All Authors: No reported disclosures

Comparison of mNGS with conventional methods for diagnosis of cryptococcal meningitis: a retrospective study

The application of metagenomic next-generation sequencing (mNGS) in the diagnosis of cryptococcal meningitis is relatively under characterized. Here, we retrospectively evaluated data from cryptococcal meningitis patients who were tested using mNGS and/or routine testing, including fungal culture, India ink staining, and cryptococcal antigen (CrAg) testing. The performance of mNGS was then assessed. Initial cerebrospinal fluid (CSF) samples were collected from 65 patients with suspected central nervous system (CNS) infection and tested using conventional tests and/or mNGS. mNGS offers a culture-independent approach, facilitating a rapid and unbiased detection of a broad spectrum of pathogens. Patients with bacterial tuberculous or viral meningitis were used as mNGS-positive controls and one autoimmune encephalitis patient was used as an mNGS-negative control. In the 45 patients diagnosed with cryptococcal meningitis, the sensitivity, specificity, positive predictive value, negative predictive value, and concordance rate of mNGS were 92%, 100%, 100%, 90.9%, and 95.6%, respectively. Compared to conventional methods, the sensitivity of mNGS was slightly lower than CrAg tests (96.7%) but higher than India ink (79.5%) and culturing (63.4%). Of the two negative mNGS cases (2/25, 8.0%), one was positive by India ink staining, culture, and CrAg testing, while the other was positive only by CrAg testing. A combination of mNGS and conventional methods enhanced the detection rate to 100%. Our study demonstrates that both CrAg and mNGS offer excellent diagnostic accuracy for cryptococcal meningitis, and utilizing both tests can enhance clinical assessment and patient management.

P-1722. Short versus Long Duration of Empiric Antifungal Use Among Patients with Sepsis or Septic Shock

Abstract Background Invasive candidiasis is associated with high mortality rates among critically ill patients. There are currently no data to guide duration of empiric antifungal therapy (EAT) when indicated in high-risk patients with sepsis. The aim of this study was to evaluate clinical outcomes associated with a short versus long course of EAT in patients with sepsis or septic shock. Methods This was a retrospective cohort study of adults admitted to the medical ICU who received &amp;gt; 2 days of EAT with fluconazole or caspofungin for sepsis or septic shock. Patients were excluded if they expired within 48 hours, had a positive fungal culture from a sterile site, were exposed to a systemic antifungal for &amp;gt; 48 hours in the 14 days preceding treatment, received prophylactic antifungals, or were neutropenic. In addition, patients who expired within 96 hours of EAT initiation were excluded from the primary outcome analysis. Patients were grouped according to receipt of EAT for ≤ 4 days (short duration) or &amp;gt; 4 days (long duration). The primary outcome was 30-day all-cause mortality. Secondary outcomes included time to mortality, hospital and ICU length of stay (LOS), vasopressor-free days over 14 days, and 30-day new proven fungal infection. Results Of 135 included patients, 79 patients received short course EAT vs. 56 patients long course (Figure 1). Baseline variables were well balanced between groups apart from older age and lower body weight observed in the short duration group (Table 1). The primary outcome, evaluable in a total of 114 patients, was observed in 33 (56.9%) patients who received a short duration and 26 (46.4%) patients who received a long duration (p=0.263; OR 1.52, 95% CI [0.73 – 3.19]). There were no significant differences in all-cause inpatient mortality or time to mortality (Figure 2). Among patients who were discharged alive, there were no significant differences in secondary outcomes with the exception of fewer vasopressor-free days in the short course group (Table 2). Conclusion Most clinical outcomes were not significantly different between patients with sepsis or septic shock who received a short versus long course of EAT. Duration of EAT is an important target for stewardship programs. This study adds to knowledge in the area, though a randomized trial may be necessary to substantiate our findings. Disclosures All Authors: No reported disclosures

Bioconversion of mustard oil cake for production of lipase, optimization and direct immobilization from solid-state fermentation extract

Fungal lipases are the leading industrial biocatalyst due to their broad applications, but high cost limits their commercial usage. The low-cost agri-residues substrates can reduce the cost of lipase production. However, the compatibility of agri-residue with fungal species, recovery process of lipase and stability of the enzyme are crucial steps. The aim of the present work was optimization of lipase production from a suitable combination of fungal culture with a locally available vegetable oilseed cake (mustard/groundnut/almond/cottonseed) in solid-state fermentation process and its direct immobilization. The enzyme produced using selected combination of Rhizopus oryzae and mustard oilseed cake was optimized by Plackett–Burman design, one-factor-at-a-time and central composite design (CCD). The highest enzyme activity of 25.08 U/gds was obtained by CCD at urea 2.11% w/w, inoculum size 1.18% v/w, and moisture content 69.99% w/w. The crude enzyme from the extract was immobilized on functionalized magnetic nanoparticles with the results of protein loading 68.88 ± 3.54 µg/mg of MNPs and activity recovery of 60.33 ± 3.03%. This study can be helpful to explore the suitability of locally available agri-residue for production of lipase and utilization of enzyme in different industrial applications.

First report of strawberry root rot caused by Monosporascus eutypoides in China.

Strawberry (Fragaria × ananassa) is an important economic crop in Hebei, China. In May 2023, root rot was observed in strawberry plantations (cultivar 'Benihoppe') in Shijiazhuang (37°57'23″N, 115°16'34″E), Hebei, China. The incidence of the disease reached up to 30% in the field. The infected plants displayed symptoms of reduced vigor and stunted growth, yellowing leaves, blackened roots, and eventual rotting. Ten diseased strawberry plants were collected for pathogen isolation. Root tissues of symptomatic plants were surface sterilized sequentially in 70% ethanol (1 min), 3.125% NaOCl (6 min), and 70% ethanol (30 s), then they were rinsed in sterile water three times (Sahu et al. 2022). The sterilized roots were then cut into 3.0 × 3.0 mm pieces and placed on potato dextrose agar (PDA) medium supplemented with chloramphenicol (100 μg/mL). After 2 to 3 days incubated at 25℃ in darkness, single colonies from the initial isolates were streaked onto PDA agar plates to obtain pure cultures. The fungal colonies appeared milky white on PDA medium and exhibited only hyphae growth. Globose perithecia were observed on V8-juice medium after 30 days of incubation, with black, smooth, spherical ascospores 13.9 to 19.1 μm in diameter (n=50). For molecular analysis, the representative isolates ME10-2 and ME10-3 from different strawberry plants were selected for the amplification of internal transcribed spacer (ITS) and elongation factor 1-alpha (EF-1α) sequences using the primer sets ITS1/ITS4 (Bellemain et al. 2010) and EF1-688F/EF1-1251R (Alves et al. 2008). The sequences were deposited in GenBank (accession no. PP990210 and PP990211 for ITS, and PQ001579 and PQ001580 for EF-1α). BLASTn analysis revealed that the sequences of ITS had above 99% (504/511 bp and 503/506 bp) similarity to Monosporascus eutypoides (MK183805.1), the sequences of EF-1α had 94% (625/661 bp and 619/656 bp) similarity to Monosporascus eutypoides (JQ958959.1). Phylogenetic analysis based on concatenated sequences of ITS and EF-1α using the Maximum Likelihood method with IQ-TREE version 2.2.3 (Nguyen et al. 2015) also indicated that the two isolates clustered with M. eutypoides. To fulfill Koch's postulates, pathogenicity tests were performed (Chen et al. 2024). Due to challenges in ascospore production, an improved wheat grain medium (wheat grain with 2% dextrose) was used for fungal culture (Sainos et al. 2006). After 14 days of inoculation at 25℃, the medium was crushed and mixed into sterilized soil with a 10% ratio by weight. Four-week-old strawberry plants were transplanted into the inoculated soil and grown in a growth chamber at 25℃ (16 h light/8 h dark). Strawberry plants transplanted into sterilized uninoculated soil served as a negative control. Three weeks post-inoculation, the inoculated strawberry plants exhibited symptoms similar to those observed in field-infected, whereas no symptoms were observed on the control plants. The pathogen was reisolated from the symptomatic plants and identified as M. eutypoides based on colony morphology on a PDA plate and ITS sequences information. M. eutypoides has been found to cause Monosporascus root rot and vine decline on muskmelon, watermelon and cucumber (Martyn and Miller 1996; Salem I. B. et al. 2013). To our knowledge, this is the first report of M. eutypoides causing root rot on strawberry. This finding suggests a potential risk of strawberry root rot when planting subsequent to cucurbit crops and assists strawberry growers in the region in adjusting their planting and disease management decisions.

Fungal infection among patients with chronic rhinosinusitis who underwent endoscopic sinus surgery

Background: Chronic rhinosinusitis (CRS) involves persistent inflammation of the nasal passages and sinuses, often attributed to Aspergillus species as a primary etiological agent. Effective treatment involves complete eradication of fungal pathogens and restoration of sinus function, commonly achieved through functional endoscopic sinus surgery (FESS). This study aimed to evaluate the clinical and radiological features, assess the prevalence and identify the mycological profile of fungal rhinosinusitis. Methods: This study was a prospective observational analysis involving a cohort of 50 patients treated at the Tirunelveli Medical College between June 2017 and July 2017. Nasal sinus tissues and secretions were collected for mycological and bacteriological analysis. Samples were processed on the same day, with fungal elements identified via KOH mount and cultured on Sabouraud dextrose agar. Bacterial cultures were grown on MacConkey agar and species identification was performed using biochemical tests. Results: Fungal culture results showed that 72% of the patients had a negative culture, while 28% had a positive culture. Among the 50 patients, 36 did not have diabetes or hypertension. All sinuses were involved in 40 patients and 24 had nasal polyps, with bilateral polyps being the most common. The most frequently isolated fungal species was Aspergillus. KOH positivity was strongly correlated with culture positivity, with 9 out of 10 KOH-positive patients also being culture-positive, while KOH negativity was associated with culture negativity in 35 out of 36 patients. Conclusions: CRS significantly affects quality of life, with fungal infections being a notable cause, especially in patients with unilateral sinus involvement. Effective treatment requires endoscopic sinus surgery, followed by antifungal therapy.

Positive sputum fungal culture, fungal sensitisation and airway microbial diversity in asthmatic children.

Sensitisation to thermotolerant fungi such as Aspergillus fumigatus and Candida albicans which can colonise the airways is associated with poor lung function in children with asthma. Dysbiosis of bacteria and fungi in the airway microbiome has been reported between health and asthma but has yet to be characterised for fungal sensitised asthmatic children. We investigated if microbial diversity of the airways is altered in fungal sensitised school-age asthmatic children. Sputum samples from children with asthma who were fungal sensitised (n=22) and non-fungal sensitised (n=17) along with children without asthma (n=15), aged 5-16 years were profiled by traditional microbiological culture, modified fungal culture, bacterial 16S and fungal ITS2 next generation sequencing. Microbiota were compared between groups using alpha/beta diversity and differential abundance analysis. Bacterial alpha diversity was significantly lower in asthma compared to disease controls and in stable compared to acute asthma. Fungal alpha and beta diversity did not change between asthma states and disease controls, but alpha diversity was significantly lower in asthma samples from patients with positive A. fumigatus culture. Children sensitised to fungi had similar microbial diversity compared to non-sensitised children. However, in children not sensitised to fungi, those with a positive airway fungal culture had significantly lower fungal alpha diversity and bacterial beta differences compared to children with negative fungal culture. Fungal sensitisation did not alter bacterial or fungal microbiota in the airways of asthmatic children. However, positive airway fungal culture was associated with significant changes in microbial diversity, particularly in non-fungal sensitised children with asthma.

The Potential of Trichoderma AA1 and AA2 as Local Indonesian Cellulolytic Inoculum in the Fermentation of Gliricidia sepium Leaf Meal

The study aimed to test the cellulolytic ability of Trichoderma AA1 and AA2 inoculum as local Indonesian isolates and their application in the fermentation of Gliricidia sepium leaf meal. Measurement of cellulolytic ability includes qualitative cellulase activity (measurement of clear zone formation around fungal colonies) and quantitative cellulase activity (measurement of the amount of reducing sugar produced by broth enzymes from fungal cultures). Experiments of Gliricidia sepium fermentation using Trichoderma AA1 and AA2 were each designed using Completely Randomized Design. The treatments used consisted of fermentation duration of 0 (control), 2, 4, and 6 days—identification of Trichoderma AA1 and AA2 species using molecular identification method. The research results show that qualitative cellulase activities in Trichoderma AA2 were significantly higher compared to Trichoderma AA1. The CZD/CD ratio of Trichoderma AA1 and AA2 were 2.39±0.11 and 1.97±0.10, respectively. The CMC-ase activity of Trichoderma AA2 was 0.332±0.038 µmol/ml/min, and this was significantly higher compared to TrichodermaAA2, which was only 0.201±0.021 µmol/ml/min. Gliricidia sepium fermentation using AA1 and AA2 inoculum showed increased nutritional value, including soluble protein content, In-vitro digestibility of dry matter, and In-vitro digestibility of organic matter in 2 and 4 days of fermentation. There was a decrease in nutritional value in 6 days of fermentation. Trichoderma AA1 and AA2 were identified as Trichoderma koningiopsis and Trichoderma asperellum, respectively. The study concluded that T. Koningiopsis AA1 and T. Asperellum AA2 were proven to have cellulolytic abilities and could be used as inoculum in Gliricidia sepium fermentation to increase its nutritional value.