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Inoculum preparation for in-vitro susceptibility testing of filamentous fungi
1Unitat de Microbiologia, Facultat de Medicina i Ciències de la Salut, Universitat Rovira i Virgili, Reus, Spain.
Abstract:
The percent transmission (T) and the equivalence in cfu/mL of two suspensions of 125 isolates of clinically important filamentous fungi containing 1-5 x 10(5) and 1-5 x 10(4) conidia/mL, adjusted with a haemocytometer, were determined. A wide variation of T values was observed, especially with the first inoculum. A good agreement was obtained between the counted and the viable propagules. The haemocytometric method is considered a useful technique for standardizing inocula.
Insights
This study found that the haemocytometer method effectively standardizes fungal inocula for clinical research. It ensures a good agreement between counted and viable fungal propagules, crucial for accurate experimental results.
Area of Science:
- Mycology
- Clinical Microbiology
- Infectious Diseases
Background:
- Accurate quantification of microbial load is essential for reproducible experimental outcomes in clinical microbiology.
- Filamentous fungi are significant human pathogens, necessitating reliable methods for inoculum preparation.
- Standardization of fungal suspensions is critical for in vitro susceptibility testing and infection models.
Purpose of the Study:
- To evaluate the utility of the haemocytometer method for standardizing inocula of filamentous fungi.
- To determine the percent transmission (T) and viable conidia/mL for fungal suspensions.
- To assess the agreement between counted and viable fungal propagules.
Main Methods:
- Determined percent transmission (T) and colony-forming units per milliliter (cfu/mL) for 125 fungal isolates.
- Prepared two concentrations of fungal suspensions: 1-5 x 10(5) and 1-5 x 10(4) conidia/mL.
- Adjusted fungal suspensions using a haemocytometer.
Main Results:
- Observed wide variations in percent transmission (T) values, particularly with higher inoculum concentrations.
- Demonstrated good agreement between haemocytometer counts and viable fungal propagules (cfu/mL).
- The haemocytometric method proved effective in achieving consistent inoculum standardization.
Conclusions:
- The haemocytometer method is a valuable and practical technique for standardizing inocula of clinically important filamentous fungi.
- This standardization enhances the reliability of experimental data in mycology and clinical microbiology.
- Accurate inoculum preparation is key to advancing research on fungal infections.