Cultivation in very different media works normally. Then you need to extract the DNA and get an ITS sequence- hope that helps

Thank you Stefan and could you tell me the special media?

Hi Rajendran,

check papers of Johan de Jonckheere e.g. 1. "De Jonckheere JF: Molecular definition and the ubiquity of species in the genus Naegleria. Protist 2004, 155(1):89-103."

That should lead you to your goals:)

naegleria fowleri has several life-cycle stages - I assume you trying to grow a flagellated organism? The media is different for each life-cycle stage - especially to induce flagella, which requires a low salt media.

My supervisor, Prof Dr Nakisah Mat Amin have conducted various research on this. You should refer to this paper

Proteinases in Naegleria Fowleri (strain NF3), a pathogenic amoeba:

a preliminary study

Plus, you can contact her for more details.

Thank you all for answering my queries and suggestions.

Not that I want to sound like some pedantic stickler but you know N. fowleri should be grown in containment level 2 facilities? Obviously by properly trained personnel as there is no treatment against infection...

Can you culture it now? I think i need your help.

Ditto what Mark can der Giezen said.  The simplest method for cultivation is on non-nutrient agar plates (water agar) coated with washed E.coli.  They form plaques as they multiply and consume the bacteria.  If you want to grow large numbers of them, you would need to get them in a liquid medium.  I recommend Mix medium (John and Howard, 1995), but they can be grown in a minimal salts medium (Page's Saline) with the washed E. coli as well.  Please work in a biocontainment hood (NOT laminar flow).  Best!

Marsha

if we want to detect Naegleria fowleri in water , what method we should use?

CLINICAL MICROBIOLOGY REVIEWS, July 2002, p. 342–354 Vol. 15, No. 3

Cultivation of Pathogenic and Opportunistic Free-Living Amebas by Frederick L. Schuster, is quite good source