Ascaridia galli in chickens: intestinal localization and comparison of methods to isolate the larvae within the first week of infection

This study was conducted to observe the localization and to compare methods for isolation of minute Ascaridia galli larvae in chicken intestine. Firstly, six 7-week-old layer pullets were orally infected with 2,000 embryonated A. galli eggs and necropsied either at 3, 5 or 7?days post infection (dpi). More than 95?% of the recovered larvae were obtained from the anterior half of the jejunoileum, suggesting this part as the initial predilection site for A. galli larvae. Secondly, the intestinal wall of one layer pullet infected with 20,000 A. galli eggs 3?days earlier was digested in pepsin?CHCl for 90?min. The initial 10?min of digestion released 51?% of the totally recovered larvae and the last 30?min of continuous digestion yielded only 5?%. This indicates that the majority of larvae were located superficially in the intestinal mucosa. Thirdly, 48 7-week-old layer pullets were infected with 500 A. galli eggs and necropsied at 3 dpi to compare three different larval isolation methods from the intestinal wall, viz., EDTA incubation, agar-gel incubation and pepsin?CHCl digestion, resulting in mean percentages of the recovered larvae: 14.4, 18.2 and 20.0?%, respectively (P?=?0.15). As conclusion, we recommended Pepsin?CHCl digestion as the method of choice for larval recovery from the intestinal wall in future population dynamics study due to high efficiency and quick and simple detection. The agar-gel method was considered to be a prerequisite for molecular and immunological investigations as the larvae were more active and fully intact.