P362 Seasonal trend of fungal flora in water of tertiary care hospital in North India

Abstract   Poster session 3, September 23, 2022, 12:30 PM - 1:30 PM Objectives The study was conducted to assess the seasonal variation of fungal flora in hospital water of a tertiary care hospital in North India. Methods A total of 200 water samples from the main reservoir, overhead and underground tanks, and taps of critical care units of the hospital were collected. The water samples were filtered by membrane filtration technique (0.22 micron) and cultured on dichloran rose-Bengal chloramphenicol agar with and without benomyl. The plates were incubated for upto 15 days and fungal colonies recovered were sub-cultured on Sabouraud Dextrose Agar and identified by phenotypic methods. Yeasts were identified by Matrix Assisted Laser Desorption Ionization Time of Flight (MALDI-TOF-MS). Results Mycelial fungi were isolated from 100% of the water samples which included Alternaria, Curvularia, Nigrospora, Penicillium, Aspergillus, Paecilomyces, Scytillidium, and Mycelia sterilia as depicted in Figure 1. Different fungi were prevailing in different water storage units like: Advance eye center— A. fumigatus, A. flavus, Fusarium solani, Alternaria spp. Rhizopus arrhizus, Ustilago spp., mycelia sterilia, Trichosporon spp.; Advance trauma Center—Cladophaialophora spp., Alternaria alternata, Penicillium, spp. A. flavus, A. nidulans, A. fumigatus, F. solani, Rhodotorula spp.: Bone marrow transplant unit— Alterneria alternata, A. niger, A. flavus, A. versicolor, Cladosporium spp., Fonsecaea pedrosoi, Fusarium spp., Nigrospora spherica, Penicillium spp., Rhodotorula spp., Trichosporon ashaii. The seasonal variation of fungal isolation is depicted in Figure 2. Isolation rate of Aspergillus species was 35% in winters, 31% in post-monsoon, 25% in summers. Isolation rate of Penicillium species was 19% in post-monsoon, 16% in winter and 15% in summers. Maximum number of dematiaceous fungi were isolated in summer season with isolation rate of 30% in summers as compared with 21.5% in post-monsoon and 19% in winters. Few yeasts isolated were Rhodotorula, Trichosporon, and Ustilago. Mucorales isolated rarely included Rhizopus, Absidia, Syncephalastrum, and Mucor species. Fungal colony forming units in the water samples ranged from 50 to 450 colony forming units/liter of water. Conclusion The distribution of fungi in hospital water showed diversity and seasonal variability. Aspergillus species were isolated in maximum number in the winter season, Penicillium species in post-monsoon season and dematiaceous fungi in the winter season. Water as a source of fungal infection in critical care units, remains a relatively neglected area. Water supply could be a source of nosocomial fungal infections. Improving the quality of water by regular testing for fungal contamination and appropriate action to reduce its burden may reduce the hospital-acquired fungal infections.

Poster session 3, September 23, 2022, 12:30 PM -1:30 PM Background: Candida auris is an emerging multidrug-resistant pathogenic yeast. The increasing frequency of C. auris outbreaks is prompting alarm worldwide. This yeast survives and spreads on contaminated medical supplies, resulting in hospital outbreaks. To learn more about the yeast's spreading behaviors and transmission, we studied its persistence and survival on a variety of medical-related surfaces under diverse environmental conditions.
Methods: A total of 104 CFU/mL solutions of four Candida species, including C. auris , C. albicans , C. parapsilosis , and C. glabrata , were inoculated onto different 2 × 2 cm sheets of cotton textile, polystyrene, paper, aluminum, glass, latex, and dried Sabouraud dextrose agar. Inoculated sheets were incubated at various temperatures and subjected to light and darkness at 1, 2, 7, 14, 30, 45, 60, and 120-day intervals. After culture of the sheets on Sabouraud dextrose agar plates, the viable CFUs of yeasts were counted.
Results: All four species remained alive on all surfaces for at least 1 week under ambient and refrigerator temperatures, darkness, and light exposure. However, only latex and polystyrene surfaces maintained viable C. auris and C.parapsilosis for a maximum of 30 days at ambient temperature and darkness. C. auris survived on dried Sabouraud dextrose agar sheets for ˃4 months.
Conclusions: Candida auris and other pathogenic yeasts can survive on a variety of medical surfaces for extended periods of time. Latex and polystyrene devices are the best medical matrices for yeast persistence. If C. auris has access to organic and nutritional components, its survival could be greatly increased. To prevent C. auris transmission, appropriate disinfection and decontamination methods should be considered.

Dona Love National Institutes of Health, Rockville, United States
Poster session 3, September 23, 2022, 12:30 PM -1:30 PM The National Institute of Allergy and Infectious Diseases ( NIAID ) funds one of the largest medical mycology research portfolios. The portfolio includes the major human fungal pathogens and covers basic fungal biology and the more translational areas of therapeutics, vaccines, and diagnostics. NIAID utilizes many granting mechanisms that are open to US and international researchers. These include investigator-initiated applications ( R01, R21, and R03s ) and targeted announcements for fungal research. Additionally, NIAID has a suite of preclinical services supporting therapeutic, diagnostic, and vaccine development. These services are free and available to investigators in academia, not-for-profit organizations, industry, or governments worldwide. The NIAID granting mechanism can be complicated. Tips and tricks for navigating the NIAID application process and preclinical services will be discussed.  Figure 2. Isolation rate of Aspergillus species was 35% in winters, 31% in post-monsoon, 25% in summers. Isolation rate of Penicillium species was 19% in post-monsoon, 16% in winter and 15% in summers. Maximum number of dematiaceous fungi were isolated in summer season with isolation rate of 30% in summers as compared with 21.5% in post-monsoon and 19% in winters. Few yeasts isolated were Rhodotorula, Trichosporon, and Ustilago . Mucorales isolated rarely included Rhizopus, Absidia, Syncephalastrum, and Mucor species. Fungal colony forming units in the water samples ranged from 50 to 450 colony forming units/liter of water.
Conclusion: The distribution of fungi in hospital water showed diversity and seasonal variability. Aspergillus species were isolated in maximum number in the winter season, Penicillium species in post-monsoon season and dematiaceous fungi in the winter season. Water as a source of fungal infection in critical care units, remains a relatively neglected area. Water supply could be a source of nosocomial fungal infections. Improving the quality of water by regular testing for fungal contamination and appropriate action to reduce its burden may reduce the hospital-acquired fungal infections.

P363 Mycoflora diversity in stored water from selected households in nairobi
Olga Mashedi 1 , Bridgit Kimani 1 , Leah Waithera 2 , Shiela Okoth 2 1 Kenya Medical Research Institute ( KEMRI ) ( 420 ) , Nairobi, Kenya 2 University of Nairobi, Nairobi, Kenya Poster session 3, September 23, 2022, 12:30 PM -1:30 PM Introduction: Water is a very vital natural resource for all life on earth as it is used widely. Water plays an important role in the world economy, the uses can be categorized into; commercial use where it is used in hostels, restaurants, offices, and other commercial activities. Fungi are ubiquitous in nature; they produce spores that are small-sized, able to stay airborne for a long duration, and transported over long distances during air dispersal. These are major sources of allergens and airway irritants that are detrimental to health. Biofilms in water distribution systems and storage containers provide favorable habitat for microorganisms as they accumulate better in solid-liquid interfaces that enable them to be embedded in the gelatinous matrix of extracellular polymers excreted by the microorganisms leading to resistance of microbes from environmental stresses. Fungi hydrophobicity and adaptability have enabled them to assemble and colonize different surfaces in domestic settings. The potential health effects caused by fungi in treated water are still not well highlighted and thus the need to carry out this study to investigate the mycoflora isolated from stored water.
Methods: This was a cross-sectional study conducted from 2019-2021, whereby 120 water samples were collected from stored containers in households in Nairobi county Kenya. The fungal strains were plated onto Sabouraud's dextrose agar ( SDA ) , Potato Dextrose Agar, and Czapecks Agar media with chloramphenicol ( 0.05 mg/mL ) ( Oxoid, U.K. ) . The plates were incubated for 7 days at 25 °C . Fungal identification was done by direct microscopy and morphological features.
Results: A total of 101 fungal species were isolated from water samples. Yeasts 48 ( 47.5% ) , Rhodotorula 34 ( 33.6% ) accounted for the unicellular fungi. Among Filamentous mycoflora, the most common isolated fungi were Aspergillus species 13 ( 12.8% ) , followed by Fusarium spp 4 ( 3.9% ) with Mucor and | Rhizopus species accounting for 1 ( 0.9% ) respectively. Yeasts species were the most common species isolated from water species and Aspergillus species were more frequently isolated from filamentous fungi.
Conclusion: Yeasts species were the most common species isolated from water species and the roles they play in biofilm needs to be further investigated.

P364 Quantitative monitoring of Aspergillus fumigatus mycelial growth by optical density
Ken Miyazawa 1 , Takashi Umeyama 1 , Yasutaka Hoshino 1 , Shogo Takatsuka 1 , Yasunori Muraosa 1 , Keietsu Abe 2 , Yoshitsugu Miyazaki 1 1 National Institute of Infectious Diseases, Tokyo, Japan 2 Tohoku University, Sendai, Japan Poster session 3, September 23, 2022, 12:30 PM -1:30 PM Objectives: Filamentous fungi generally form hyphal pellets in liquid culture. This property prevents filamentous fungi from applying the growth monitoring methods used for unicellular organisms such as yeast and bacteria. We have analyzed the biological functions of cell wall polysaccharide α-1,3-glucan and extracellular polysaccharide galactosaminogalactan ( GAG ) in Aspergillus species, and revealed that both α-1,3-glucan and GAG contribute to the pellet formation. Here we constructed the