title: Establishing mono-eukaryotic Histomonas meleagridis cultures from in vivo infection contaminated with Tetratrichomonas gallinarum and Blastocystis spp.
abstract: SUMMARY The necessity to easily establish Histomonas meleagridis cultures has been underlined extensively by many researchers in order to gain more insights in the biology of H. meleagridis. In addition the occurrence of different protozoa in the caeca of birds impedes, however, the isolation and propagation of H. meleagridis from field outbreaks. Therefore, in a kinetic study using transmission electron microscopy the deleterious effects of adventitious protozoa including Tetratrichomonas gallinarum and Blastocystis spp. on cultured H. meleagridis were examined. To overcome this issue, an easy and successful approach to establish the mono-eukaryotic H. meleagridis culture free of other host's protozoa is proposed. At 10 days post infection, liver lesions of H. meleagridis-infected birds were isolated and inoculated into culture media pre-incubated with caecal bacteria. After 48 h of incubation, presence of H. meleagridis in the cultures was confirmed through morphological evaluation. Additionally, TEM examination and analysis by PCR amplification of the small subunit rRNA gene could exclude the co-cultivation of T. gallinarum and Blastocystis spp. Furthermore, after successful propagation and maintenance of the cultured H. meleagridis, its pathogenicity was affirmed in an infection experiment in turkeys.
authors.affiliation: Laboratory of Livestock Physiology, Immunology and Genetics, KULeuven, Kasteelpark Arenberg 30, 3001 Heverlee, Belgium.
authors.forename: Anh Dao Nguyen
authors.initials: AD
authors.lastname: Pham
authors.affiliation: Electron Microscopy Unit, Veterinary and Agrochemical Research Centre, CODA-CERVA, Groeselenberg 99, 1180 Ukkel, Belgium.
authors.forename: Jan
authors.initials: J
authors.lastname: Mast
authors.affiliation: Poulpharm, Ninovestraat 7, 9420 Erpe-Mere, Belgium.
authors.forename: Jeroen Koen
authors.initials: JK
authors.lastname: DE Gussem
authors.affiliation: Department of Poultry Science, University of Georgia, Athens, Georgia 30602, USA.
authors.forename: Larry R
authors.initials: LR
authors.lastname: McDougald
authors.affiliation: Laboratory of Livestock Physiology, Immunology and Genetics, KULeuven, Kasteelpark Arenberg 30, 3001 Heverlee, Belgium.
authors.forename: Bruno Maria
authors.initials: BM
authors.lastname: Goddeeris
mesh_terms: D000818:Animals; D016844:Blastocystis / Q000254:growth & development / Q000502:physiology* / Q000648:ultrastructure; D046508:Culture Techniques / Q000592:standards; D008297:Male; D046529:Microscopy, Electron, Transmission; D016133:Polymerase Chain Reaction; D011201:Poultry Diseases / Q000469:parasitology*; D011529:Protozoan Infections, Animal / Q000469:parasitology*; D016834:Trichomonadida / Q000235:genetics / Q000254:growth & development / Q000472:pathogenicity / Q000502:physiology* / Q000648:ultrastructure; D014422:Turkeys