Tap water Five hundred millilitre o

Tap water
Five hundred millilitre of tap water from each of 50 taps was filtered through sterile 0.45 μm membrane filter within 2 hours of collection.[4] Filters were then inverted onto 1.5% non-nutrient agar (NNA) plates with an overlay of E. coli ATCC 25922. All plates were sealed and incubated at 37°C. They were examined daily for growth of amoebae under 10 × magnification of light microscope for up to 2 weeks. The FLA were confirmed by their cyst and trophozoite morphology. A block was cut and placed onto fresh NNA plate overlaid with E. coli and incubated at 30°C for subculture to obtain growth for molecular study.