Gastrothylax crumenifer: ultrastructure and histopathology study of in vitro trematodicidal effect of Microlepia speluncae (L.) Moore

An in vitro incubation study was carried out to determine the efficacy of Microlepia speluncae (MS) fronds' extract on the worm motility and viability, morphology and histopathological changes in/of Gastrothylax crumenifer. The methanolic extract of MS fronds was used for in vitro study at 1, 2, 3, 4 and 5 mg/ml concentration as test extracts. Oxyclozanide (OXY) at 1% (250 mg/25 ml) as standard control and Hedon-Fleig (H-F) salt solution as negative control. The solutions (25 ml), with 25 amphistomes each, distributed to 7 petriplates (90 mm diameter) and placed in an incubator with 5% CO 2 at 37°C. The motility of control and treated flukes was observed at regular time intervals, 0, 10, 15, 30 and 60 min respectively with specific score 3, 2, 1, 0 for motility. Relative Motility assay and LC 50 were determined by probit regression analysis. In vitro incubation study revealed death of all trematodes, lethal at 10 min incubation time at 5 mg/ml concentration, indicated RM value is 0, very much effective than Oxyclozanide, RM value was 0. 05. The LC 50 was determined as 3.666. The worms subjected to in vitro trials were studied under Scanning Electron Microscope and Light Microscope for significant morphological changes.