Table 1.
Egg counts of C. sinensis daily produced by one adult worm in different media during the cultivation.
Table 2.
Measurements of eggs obtained from different media after cultivation (n = 15).
Figure 1.
Light microscopic changes of C. sinensis eggs in different solutions and media during cultivation.
Morphological view of eggs from day 1 to day 90 arranged from top to bottom. As the worm survived up to 60 days in 1× Locke’s solution there was no data for day 90. All the eggs showed typical morphology at day 1, however, at day 90 eggs showed atypical shape, shorter length, prominent abopercular knob along with interior dark granules (×400). Scale bar: 10 µm.
Figure 2.
Deformed C. sinensis eggs observed after 90 days of incubation in different broth media.
(A) Normal egg. (B) Egg containing deformed granules. (C) Egg containing dark granules both at opercular and abopercular ends. (D) Dark granules fragmented in to smaller granules. (E) Fragmented dark granules covered whole interior part of the egg. (F) Degraded dark granules in side the egg with prominent abopercular knob. (G) Egg with deformed shape and degraded miracidium. (H) Extremely deformed egg with degraded larval products. Original magnification ×400. Scale bar: 10 µm.
Figure 3.
Surface ultrastructure of C. sinensis eggs by scanning electron microscopy.
Eggs after 60 days of incubation in 1× Locke’s solution (A) and 90 days of incubation in RPMI-1640 (B), DMEM (C) and IMDM (D) culture media (×2000). Arrow head showed smoothness on the surface or loss of wrinkles. Scale bar: 10 µm.
Figure 4.
Abopercular knob of normal and cultured C. sinensis eggs.
(A) Light microscopy showed an inconspicuous abopercular knob in a normal egg (×1000). (B) Very prominent abopercular knob in cultured worm’s egg (×1000). (C–D) SEM images of normal and cultured worm’s eggs respectively (×2000). (E) Magnified posterior portion of normal egg (×4000). (F) Posterior portion of cultured worm’s egg with distinct abopercular knob (×4000). Scale bar: 10 µm for A–D and 5 µm for E–F.
Table 3.
Proportion of viable eggs in different solution and media during cultivation determined by trypan blue (0.4%).