Eimeria tenella is recognized worldwide as a significant pathogen in the poultry industry. However, the lack of a method for isolating developing schizonts has hindered the use of transcriptome analyses to discover novel and developmentally regulated genes. In the present study, we characterised the long-term successive development of E. tenella in infected domestic fowl caeca and assessed the utility of laser microdissection (LMD) for the isolation of schizont RNA. Developmental stages were synchronous, including those of the first-, second-, and third-generation schizonts and gametocytes; the second-generation schizont stage was associated with the pathogenicity of the disease, such as severe bloody diarrhoea and eventual death. Using LMD, the mature second-generation schizonts were successfully excised from the lamina propria, and non-degraded RNA was purified from the schizonts. Eimeria tenella-specific genes were amplified by reverse transcription polymerase chain reaction (RT-PCR). These results augment our understanding of the E. tenella life cycle, and reveal the potentially utility of LMD as a tool for gene expression analyses of the intracellular stages of E. tenella.
(Bacterial and Parasitic Disease Research Division)
References:
Matsubayashi M. et al. (2012) Parasitology 139(12):1553-1561
