The fine structure of the cuticle of the infective larva and of the adult male and female was studied in Mastophorus muris, a stomach parasite of rats. The infective larva develops in the hemocoel of the insect intermediate host where it lays coiled in a gelatinous cyst with its ventral surface outwards. In transverse sections of uncoiled larvae the cuticle is 2.4 uAm thick and consists of a cortex in which a narrow band of striations is visible, a median layer, and one layer of basal fibers. In coiled specimens the ventral cuticle of the larva is stretched and thin, whereas the dorsal cuticle is thicker, folded, and appears compressed. The cuticle of the adult worms is 17-74 Arm thick and consists of an outer fuzzy coat, an external trilaminar structure, a granular external cortex, and an internal cortex containing striated fibers with a periodicity of 200 nm. The median layer consists of homogeneous, electron-dense material in which less electron-dense material is seen. Three sublayers of fibers comprise the basal layer. The innermost basal lamella consists of striated fibers with a 140-nm period. The ultrastructure of nematode cuticle has been reviewed by Lee (1966b, 1972), Bird and Bird (1969), and Bird (1971). With the exception of several species of filariae, representatives of the suborder Spirurina have been studied only by light microscopy (Bogoyavlenski, 1961). In the present study the cuticular ultrastructure of adult Mastophorus muris, parasitic in the stomach of rats, and of its infective third stage larva, parasitic in the hemocoel of the cockroach, is described. MATERIALS AND METHODS Mastophorus muris, isolated from city rats (Wertheim, 1963), was maintained in the outbred Hebrew University Sabra strain of rats and in Blatella germanica. Roaches, fasted for 48 hr, were infected with larvated eggs mixed with powdered wet oat flakes. The rats, 80-100 g, were infected by gavage with 15 gelatinous cysts containing 3rd stage larvae, dissected from the roaches 4-5 weeks postinfection. For this study adult worms were removed from Received for publication 6 September 1977. * On Fellowship Leave from LaGuardia Community College, City University of New York, Long Island City, New York. the stomach of ether-killed rats, infected 60 days previously. The 3rd stage larvae were freed from the gelatinous cysts by treatment with artificial gastric juice for 10 min at 37 C. Worms were rinsed in saline and cut into appropriately sized cylinders. Tissue was fixed for 1-2 hr in ice-cold 2.5% glutaraldehyde in either phosphate or cacodylate buffer pH 7.4. Worms were then rinsed in 3 changes of cold buffer for 1 hr. Postfixation was carried out for 1 hr with ice-cold 1% osmium tetroxide in one of the above mentioned buffers. The tissue was rinsed in distilled water and in 2% uranyl acetate in distilled water, dehydrated in an ethanol series, transferred to propylene oxide, and left overnight in a 1:1 mixture of propylene oxide-Epon. Following 2 hr in fresh Epon in vacuo (Bird, 1971), the tissue was embed ed in fresh Epon in flat silicone rubber molds, oriented, and polymerized at 60 C for 48 hr. Blocks were sectioned with glass knives on a LKB Ultratome. Thick (0.5-1.0 tum) sections were stained with 1% toluidine blue in 0.5% sodium borate at 60 C. Selected areas were thin sectioned and picked up on naked or formvarcarbon coated grids, stained with uranyl acetate and lead citrate, and examined with a Philips 300 electron microscope at 60 kV. Worms for scanning electron microscopy were prepared according to the method descibed elsewhere (Wertheim and Chabaud, 1977). FIGURES 1, 2. Third stage larva of Mastophorus muris. 1. Electron micrograph of a transverse section from the middle of uncoiled larva. The striated band is visible near the interior border of the cortex. Cortex (C), median (M), and basal (B) layers. X 53,000. 2. Transverse section of the lateral alae of larva. Body wall layers are retained, cortex (C), median (M), and basal (B), and an electron-dense granular deposit (G) is visible between the cortex (C) and median (M) layers. Cyst debris is visible on external surface of the cuticle. X 20,000.
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