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Clinically relevant fungi in water and on surfaces in an indoor swimming pool facility
Published in Yuli Ekowati, Protection of Public Health from Microbial and Chemical Hazards in Swimming Pool Environments, 2019
Fungal DNA from the isolated colonies that were grown on OA as described above was extracted using Chelex® 100 (Bio-Rad Laboratories, USA). The extraction was done by harvesting 5–10 mm3 of mycelium using an inoculation needle and subsequently putting the mycelium in sterile reaction tubes together with 5–10 acid-washed glass beads (1.5–2 mm). The tubes with fungal material were frozen in liquid nitrogen for 3–5 minutes after which a bead beater was used to disrupt the cells for 10 seconds at 30 beats/sec. Both freezing and bead beating were repeated 2 times. Afterwards, 100 µL of 5% Chelex resin and 10 µL Proteinase K (20 mg/mL) were added to the lysed samples. The tubes were incubated for 30 min at 56 °C followed by 10 min at 95 °C using dry block heaters and then centrifuged for 1 min at 12,000 rpm to settle the beads and resin. The supernatant was transferred to a new reaction tube. The centrifugation was repeated 2–3 times until all resin was removed. The extracted DNA was stored at -20 °C until further analysis.
Evaluation of the effectiveness of two Iranian su-strains of Metarhizium anisopliae (ascomycota: hypocreales) on the mortality rate of American cockroach
Published in International Journal of Environmental Health Research, 2023
Hamed Ramezani Awal Riabi, Mehran Ghazavi
The Nour (DEMI002) and Saravan (DEMI001) isolates of M. anisopliae were obtained from the Agricultural Entomology Research Department of the Iranian Plant Pathology Research Institute. The study employed the single-spore method using the PDA (170 g potato, 20 g dextrose, 18 g agar, per 1000 mL) in a petri dish (80 mm) to purify the fungus. Environments were maintained at 25 ± 2°C for two weeks. Sporulation of the isolates was determined using slide and microscopic magnification of 40 × . The conidia were harvested by scraping the surface of a 16-day-old culture gently with an inoculation needle and were suspended in 10 cc of distilled water mixed with 2 drops of 0.01% (v/v) Tween-80. The suspension then being put in a centrifuge for 15 seconds at 1000 rpm. Finally, the prepared suspension was passed through 3 layers of sterile gas, and the spores were purified.
Rapid degradation of FOG discharged from food industry wastewater by lipolytic fungi as a bioaugmentation application
Published in Environmental Technology, 2018
Ayoma Witharana, Jagath Manatunge, Niranjanie Ratnayake, Chandrika M. Nanayakkara, Mahesh Jayaweera
A fungal spore suspension was obtained by growing a pure culture of particular isolate on minimal salt agar and incubating at 30°C for seven days. A total volume of 10 mL sterile distilled water was spread in aliquots on a culture plate and the fungal colony surface was gently scraped using an inoculation needle. The cultures were filtered through Whatman® No. 42 filter paper (2.5 µm) into a sterile container. An aqueous spore suspension having approximately 1 × 107 spores per 1 mL was obtained using a hemocytometer. Microscopic and macromorphological characteristics of isolates have been used to identify fungi isolates upto the genus level.