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Entamoeba dispar : Cultivation with Sterilized Crithidia fasciculata 1
Author(s) -
Kobayashi Seiki,
Imai Eiko,
Tachibana Hiroshi,
Fujiwara Tatsushi,
Takeuchi Tsutomu
Publication year - 1998
Publication title -
journal of eukaryotic microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.067
H-Index - 77
eISSN - 1550-7408
pISSN - 1066-5234
DOI - 10.1111/j.1550-7408.1998.tb04516.x
Subject(s) - crithidia fasciculata , dispar , biology , entamoeba , entamoeba histolytica , microbiology and biotechnology , crithidia , protozoa , virology
Four isolates of Entamoeba dispar identified by their hexokinase and phosphoglucomutase isoenzyme profile and by their failure to react with Entamoeba histolytica ‐specific monoclonal antibody (4G6) could be grown in either Diamond's BI‐S‐33 medium, newly developed BCSI‐S (Biosate cysteine starch iron‐serum) medium, or casein‐free YI‐S medium in the presence of Crithidia fasciculata (ReF‐1:PRR) sterilized by heating 56° C for 30 min and subsequent incubation with 1% hydrogen peroxide for 24 hours at 4° C. After the cultures were maintained for over 50 passages, the amebae were identified as E. dispar by isoenzyme analysis, polymerase chain reaction with E. histolytica ‐ and E. dispar ‐specific primers, i.e. p11 plus p12 and p13 plus p14, respectively, and by negative reactivity with monoclonal antibody 4G6. The flagellates added to the culture were judged to be metabolically inactive based on the results of nuclear magnetic resonance spectroscopy, electron microscopy, and polarographic analysis. All of these findings suggest that E. dispar can grow in vitro with metabolically inactive C. fasciculata as a culture associate.