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Establishing mono-eukaryotic Histomonas meleagridis cultures from in vivo infection contaminated with Tetratrichomonas gallinarum and Blastocystis spp.

Published online by Cambridge University Press:  21 June 2013

ANH DAO NGUYEN PHAM ,
JAN MAST ,
JEROEN KOEN DE GUSSEM ,
LARRY R. MCDOUGALD  and
BRUNO MARIA GODDEERIS
ANH DAO NGUYEN PHAM*
Affiliation:
Laboratory of Livestock Physiology, Immunology and Genetics, KULeuven, Kasteelpark Arenberg 30, 3001 Heverlee, Belgium
JAN MAST
Affiliation:
Electron Microscopy Unit, Veterinary and Agrochemical Research Centre, CODA-CERVA, Groeselenberg 99, 1180 Ukkel, Belgium
JEROEN KOEN DE GUSSEM
Affiliation:
Poulpharm, Ninovestraat 7, 9420 Erpe-Mere, Belgium
LARRY R. MCDOUGALD
Affiliation:
Department of Poultry Science, University of Georgia, Athens, Georgia 30602, USA
BRUNO MARIA GODDEERIS
Affiliation:
Laboratory of Livestock Physiology, Immunology and Genetics, KULeuven, Kasteelpark Arenberg 30, 3001 Heverlee, Belgium
*
*Corresponding author: Laboratory of Livestock Physiology, Immunology and Genetics, KULeuven, Kasteelpark Arenberg 30, 3001 Heverlee, Belgium. E-mail: [email protected]
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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.

Keywords

Histomonas meleagridisTetratrichomonas gallinarumBlastocystis spp.liverculturepathogenicitypoultryblackhead diseasehistomonosis
Type
Research Article
Information
Parasitology , Volume 140 , Issue 10 , September 2013 , pp. 1266 - 1274
Copyright
Copyright © Cambridge University Press 2013 

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