Angiostrongylus cantonensis: tegumental and hypodermic alterations of the fourth-stage larvae following administration of tribendimidine in vivo and in vitro.

Angiostrongylus cantonensis is a parasitic pathogen whose forth-stage larvae (L4) parasitize in the central nervous system (CNS) of the human cause severe eosinophilic encephalitis or meningoencephalitis. Previous study indicated an impressive anthelmintic efficacy of tribendimidine (TBD) against CNS parasitized L4 of A. cantonensis. Tegument of the larvae is the first physical barrier to protect them from attack by the host immune system. In the present study, tegumental and hypodermic alterations were observed by scanning electron microscopy and transmission electron microscopy after administration of TBD. During treatment of TBD in vivo, L4 presented wizened side sensor, disappearance of mastoids and longitudinal grain, prominent surface coat, heterogeneous tegumental layers, incompact hypodermic cell junctions, blurred myotube, and small scale of vacuole in a basal layer. After incubation with TBD in vitro, L4 exhibited a swollen side sensor and mastoids disappearance in head end. Abundant tegumental blebs and obvious deformation of both cross-grain and longitudinal grain were detected on the surface, and shrinkage of all tegumental layers, chaotic cell junction, turbid muscle cell, disappearance of myotubes, and vacuole-like changes were visible under the electron microscope. The results implied the potential mechanism of the anthelmintic effect of tribendimidine against L4 of A. cantonensis by direct damages to tegumental and hypodermic.

Angiostrongylus cantonensis: scanning electron microscopic observations on the cuticle of moulting larvae.

Angiostrongylus cantonensis is a parasitic nematode that needs to develop in different hosts in different larval stages. Freshwater snails, such as Pomacea canaliculata, are the intermediate host, and rats are the definitive host. Periodic shedding of the cuticle (moulting) is an important biological process for the survival and development of the parasite in the intermediate and definitive hosts. However, there are few studies on the cuticle alterations between different stages of this parasite. In this study, we observed the ultrastructural appearance and changes of the cuticle of the 2nd/3rd stage larvae (L2/L3) and the 3rd/4th stage larvae (L3/L4) using a scanning electron microscope. We also first divided L2/L3 into late L2 and early L3. The late L2 lacked alae, but possessed a pull-chain-like fissure. Irregular alignment of spherical particles on the cuticle were noted compared to the L3. Alae appeared in the early L3. The old cuticle turned into a thin film-like structure which adhered to the new cuticle, and spherical particles were seen regularly arranged on the surface of this structure. Regular rectangular cavities were found on the surface of L3/L4. The caudal structure of L3/L4 was much larger than that of L3, but caudal inflation, such as seen in L4, was not observed. These results are the first to reveal the ultrastructural changes of the cuticle of A. cantonensis before and after moulting of L2/L3 and L3/L4.