The L3 to L4 molt of Brugia malayi: real time visualization by video microscopy

Brugia malayi and other filarial parasites have been studied in great detail, especially in the context of human disease. In common with other nematodes, these organisms molt 4 times in their life cycles, but details of this process have not been described. We have recently developed an in vitro culture system that supports the L3 to L4 molt at high efficiency. This has permitted us to visualize, for the first time, details of this molt using real-time video microscopy. Molting is preceded by a phase of altered motility during which the larva exhibits contractile, coiling movements. The earliest evidence of ecdysis is a clearing at one end, more frequently caudal, caused by the larva retracting from that end. A cleavage develops in the cuticle near the head end, forming a rostral cap, which is continuous with the pharyngeal cuticle. Simultaneously, it retracts out of the cuticle using coiling and writhing movements. This process takes 5 to 10 min. Finally, it retracts out of the cap and extrudes the pharyngeal cuticle. Detachment of the pharyngeal cuticle is the final event in the process and continues up to an hour after the rest of the cuticle has been shed.