Detection of Histomonas meleagridis in turkeys cecal droppings by PCR amplification of the small subunit ribosomal DNA sequence

Histomonas meleagridis is a protozoan parasite that may cause histomoniasis, a disease of gallinaceous fowl characterized by necrotic typhlitis, hepatitis and high mortality. Diagnosis of this disease is based on direct identification or on cultivation of the parasite. With the aim of developing more sensitive, rapid and useful tools for parasite detection, PCR that amplified a DNA target of 209 pb of the 18S rRNA gene was designed to detect the genome of H. meleagridis and to differentiate it from the genome of Tetratrichomonas gallinarum, another common protozoan parasite of fowl. The sensitivity of the test was evaluated using serial diluted samples of cultured H. meleagridis and showed positive amplification for concentrations comprised between 10 and 10(-1)parasites/ml of culture. The sensitivity for cecal droppings samples was assessed using spiked material and was comprised between 3 x 10(3) and 3 x 10(5)parasites/ml of stool. The reliability of the PCR for the detection of Histomonas infection was also evaluated by experimental infection of turkeys. Results of the PCR appeared to be in agreement with the development of the clinical signs and of the cecal lesions. The PCR developed in this study may be a useful tool in the detection and identification of H. meleagridis for rapid, routine screening as a supplement to direct identification or cultivation of the parasite.