Gastrothylax crumenifer: Ultrastructure and histopathology study of in vitro trematodicidal action of Marattia fraxinea (Sm.)

Abstract

Background: Trematodicidal action of Marattia fraxinea (Sm.) rhizome, extracts against adult Gastrothylax crumenifer and its morphological changes under light and scanning electron microscope was studied in this work. This plant according to folk medicine has been reported to be used as antioxidant, antimicrobial and anthelmintic against ankylostomiasis. Methods: Preliminary screening involved the qualitative methods to detect the presence tannins, saponins, quinones, terpenoids, steroids, flavonoids, phenol, alkaloids, glycosides, cardiac glycosides, coumarins, anthocyanin and betacyanin. Total phenol, total terpenoids, total tannin and total flavonoids were quantitatively estimated. Total phenolic content was estimated by Folin-Ciocalteau method. Total flavonoid content was determined by the aluminium chloride colorimetric method. In-vitro incubation study of Marattia fraxinea Sm. extracts against Gastrothylax crumenifer were performed using 25 ml of Hedon-Fleig (H-F) salt solution containing various concentrations (1, 2, 3, 4 and 5 mg/ml) respectively as test extracts, positive control Oxyclozanide @ 1% (250 mg/25 ml) and negative control H-F salt solution, distributed to 7-Petri plates in an incubator with 5% CO 2 at 37°C. Twenty five amphistomes were incubated and the motility of control and test group was observed under dissection microscope at a regular time interval of (0, 10, 15, 30 and 60) min respectively. The motility response of the parasites was categorized with specific score 3, 2, 1, 0 respectively. Relative Motility (RM) value and Lethal Concentration 50 (LC 50 ) were determined by probit regression analysis. Ultrastructure and histopathological changes on morphology of G.crumenifer were interpreted. The bioactive compounds were analysed using gas chromatography-mass spectrometry (GC-MS) instrument. Results: Phytochemical analysis of M.fraxinea (Sm.) revealed the presence of secondary metabolites tannins, saponins, quinones, terpenoids, steroids, flavonoids, phenol, alkaloids, cardiac glycosides, coumarins, and betacyanin. Total terpenoid, total phenol, total tannin and total flavonoid content were found to be 16.00 ± 0.85 mg/g. 5.40 ± 0.71) mg GAE/g, 7.90 ± 0.24 mg TAE/g and 1.25 ± 0.41 mg QE/g respectively. GC-MS analysis showed presence of PHYTOL, a diterpenoids, fern-8-ene, a triterpenoids shown to have potent anthelmintic property. In vitro incubation study revealed death of all trematodes, lethal at 60 min incubation time at 5 mg/ml concentration, indicated Relative Motility (RM) value is 0, least effective compared to positive control Oxyclozanide, where RM value is 0 at 10 min. Lethal concentration 50 (LC 50 ) was found to be 4.873 mg/ml. Confirmative study on gross morphology, histopathology, and ultra-structural changes showed considerable effect on suckers, teguments and internal organs and were dose dependent. Conclusion: It is indicative that presence of terpenoids, flavonoids, tannins and phenolics which are contributing factors for anthelmintic activity, producing various ...

Untitled

“…The present results are similar with the previous study that the researchers were observed disruptions, disarrangement, and detachment of tegument from the basal membrane, eruption, blebs, erosions and vacuoles in the tegumental structure, swelled deformed body, retracted sucker, morphological and structural changes caused by Balanites aegyptica, Plumbago indica, Dregea volubilis and the oil of Nigella sativa treated on adult Paramphistomum microbothrium, Paramphistomum cervi, Gastrothylax crumenifer, Paramphistomum explanatum and Fasciola gigantic 8,10,25,28,29 . In the present study synthetic drug albendazole also shows mild anthelminthic activity and serves few damages and vacuoles on the tegument of Cotylophoron cotylophorum.…”

Anthelmintic Activity of Ocimum sanctum Linn. Leaves Ethanol Extract Against Fasciola gigantica in vitro

“…leaves ethanol extract has been proved to have phytochemical constituent that beneficial to be anthelmintic. Anthelmintic is the kind of drug which has harming effects to the helminths by affecting the vital activities, transmission of neuromuscular, ion exchange or to the tegument system that caused the helminth to be disintegrated and become paralysis or dead, therefore it will be expelled from the host's body (Nilani et al, 2012 andRajesh et al, 2017).…”

Untitled

“…[60] In vitro anthelmintic activity of plants have been studied on trematode parasites liver fluke Fasciola gigantica [61,62] tegument of G. crumenifer and on amphistome Orthocoelium scoliocoelium, F. gigantica. G. crumenifer [63][64][65][66][67] and intestinal helminthes [Table 1]. [68]…”