Method to count fungi
Technically, how to count fungal spores
1. Prepare suspensions of the analysed product, containing fungal spores in distilled water as needed to have optimal density of particles in a view field, it is chosen experimentally. The suspension of the tested product could be filtered through cheesecloth.
2. Prepare one Neubauer’s haemocytometer: clean all surfaces of the hemacytometer and cover glass, take care to ensure that all surfaces are completely dry using lint-free tissue, center the coverslip on the hemacytometer.
3. Put 9 µl of the cell suspension into one of two counting chambers of Neubauer’s haemocytometer. Use a clean pipette tip. Be sure that the suspension is thoroughly, but gently, mixed before drawing the samples. Fill the chambers slowly and steadily. Avoid injecting bubbles into the chambers. Do not overfill or under-fill the chamber.
4. Count the fungal spores under light microscope. Don't count talc particles of the product in the suspension. Count all of the fungal spores in four corner squares of the chamber. Counting criteria: Don't count the spores touching the top or left borders and don't count the spores touching the bottom or right borders of squares.
5. Calculate the total number of the fungal spores counted in 4 squares. Calculate concentration of fungal spores in the suspension using the equation: spores/ml = n x 10⁴, where n is the average number of spores counted in 4 squares of the chamber.
6. Recalculate the concentration of the fungal spores in the product according dilution of suspension.
References:
1. M. Kamaruzzaman, M. Delwar Hossain, and I. Hossain. Antifungal and Morphological Assay of Selective Trichoderma Isolates Against Soil Borne Plant Pathogenic Fungi. International Journal of Innovation and Applied Studies ISSN 2028-9324 Vol. 16 No. 2 Jun. 2016, pp. 409-417.
2. Mukhtar I., Hannan A., Atiq M., Nawaz A. Impact of Trichoderma species on Seed Germination in Soybean//Pak. J. Phytopathol. 2012. Vol. 24. No. 2. P. 159-162.
3. Sonja TANČIĆ ŽIVANOV1*, Siniša JOCIĆ1, Vladimir MIKLIČ EFFICACY OF TRICHODERMA SPP. ISOLATES AGAINST SCLEROTINIA SCLEROTIORUM ON SUNFLOWER SEEDLINGS 19th International Sunflower Conference, Edirne, Turkey, 2016.