Clonal cultures of Histomonas meleagridis, Tetratrichomonas gallinarum and a Blastocystis sp. established through micromanipulation.
Clonal cultures of Histomonas meleagridis, Tetratrichomonas gallinarum and a Blastocystis sp. were established for the first time. Single microbes were successfully isolated from a mixture of micro-organisms obtained from caecal contents of turkeys, using a micromanipulation approach. The cloned parasites were propagated in vitro and maintained through continuous passages multiplying to high numbers. Identification of the protists was done by morphological investigation identifying various forms of each parasite. PCR and partial sequencing of the small subunit rRNA were used to confirm clonality and to determine the relationship of the cloned parasites with known protozoan parasites. The clonal cultures established by this technique will be useful to gain more insight into the biological repertoire of the organisms. In addition, refined infection experiments in different poultry species can now be performed to elucidate the pathological pathways of the respective protozoa.
Hess
M
M
Clinic for Avian, Reptile and Fish Medicine, University of Veterinary Medicine, Vienna, Veterinärplatz 1, 1210 Vienna, Austria. michael.hess@vu-wien.ac.at
Kolbe
T
T
Grabensteiner
E
E
Prosl
H
H
D000818:Animals; D002988:Cloaca / Q000469:parasitology; D019976:Cloning, Organism / Q000379:methods*; D015169:Colony Count, Microbial; D003470:Culture Media; D056890:Eukaryota / Q000145:classification / Q000235:genetics / Q000254:growth & development*; D005243:Feces / Q000469:parasitology; D008846:Micromanipulation; D008853:Microscopy; D016133:Polymerase Chain Reaction; D011201:Poultry Diseases / Q000469:parasitology; D011529:Protozoan Infections, Animal / Q000469:parasitology; D016053:RNA, Protozoan / Q000235:genetics; D012335:RNA, Ribosomal / Q000235:genetics; D012692:Serial Passage; D013045:Species Specificity; D014422:Turkeys / Q000469:parasitology
D003470:Culture Media; D016053:RNA, Protozoan; D012335:RNA, Ribosomal