Evaluation of in vitro anthelmintic activity of crude extract and synthesized green silver nanoparticles of the leaves of Mammea americana L

Abstract

One of the most severe parasite infections to ever plague animals, paramphistomosis is particularly devastating in tropical and subtropical areas and causes significant economic losses to the world's food industry and poor animal health. In the present study potency of the crude extract and green synthesized silver nanoparticles from fresh and mature Mammea americana L. leaves was evaluated on trematode Paramphistomum cervi, collected from the infected rumen of the cow (Bos indicus) in 0.9% phosphate-buffered saline (PBS). The crude extract at five dose levels (1 mL, 2 mL, 3 mL 4 mL, and 5 ml), and silver nanoparticles (AgNps) at the five dose levels (10 mg/mL, 30 mg/mL, 50 mg/mL, 70 and 90 mg/mL) were applied. The synthesized AgNPs were characterized using UV-VIS spectrophotometry, X-ray diffraction (XRD), field emission-scanning electron microscopy (FE-SEM), transmission electron microscopy (TEM), and Fourier Transform Infrared Spectroscopy (FTIR) analysis. Characterization studies of AgNPs revealed the synthesis of stable, crystalline AgNPs measuring an average size of 20-50 nm. The efficacy was evaluated based on the spontaneous movement reduction and death of P. cervi. Maximum anthelmintic activity was found at the dose of 5 mL of crude extract and 90 mg/mL of AgNPs of M. americana leaves. The scanning electron micrograph study of treated helminths of crude and AgNps revealed conspicuous distortion on the surface architecture. It also showed shrinkage of the whole body and severe damage on both suckers (anterior and posterior). AgNPs caused more shredded and torn tegumental surfaces with loss of tegumental papillae compared to untreated. Thus, this study confirmed that the M. americana leaves extract and synthesized AgNPs can be a potential alternative herbal drug in traditional folklore medicine against serious helminth infections in our livestock industry.

Keywords: Anthelmintic activity; Mammea americana; Paramphistomum cervi; Scanning electron microscopy; Synthesized silver nanoparticles.

Fig. 1

a initial colour of leaf aqueous extract, b colour changes after addition of AgNO₃ solution

Fig. 2

Characteristic surface plasmon absorption spectrum (λmax = 420 nm) of the dilute dispersed silver nanoparticle solution of M. americana

Fig. 3

(a, b) Field Emission Scanning Electron Microscope (FESEM) image of synthesized AgNPs from the leaves of M. americana

Fig. 4

(a, b, c) TEM images of synthesized silver nanoparticles from leaves of M. Americana, at 100 nm scale a, 20 nm scale b and c shows enlarged view of the polyhedral shape of silver nanoparticles

Fig. 5

X-Ray diffraction (XRD) pattern of silver nanoparticles synthesized from the leaves of M. americana

Fig. 6

FTIR spectra of AgNPs synthesized by mature leaves extract of M. americana

Fig. 7

Scanning Electron Micrograph (SEM) of untreated adult P. cervi (a, b, c and d) [Here, AS (anterior sucker), P (papillae), GP (gonopore), TG (tegument), and AC (acetabulum)]

Fig. 8

Scanning Electron Micrograph (SEM) of the adult P. cervi treated with crude extract of M. americana leaves (a, b, and c) compared with untreated P. cervi (7a, 7b and 7c) [Here, AS (anterior sucker), P (papillae), GP (gonopore), TG (tegument), and AC (acetabulum)]

Fig. 9

Scanning Electron Micrograph (SEM) of the adult P. cervi treated with synthesized AgNPs of M. americana leaves (a, b, and c) compared with untreated P. cervi (7a, 7b and 7c) [Here, AS (anterior sucker), P (papillae), GP (gonopore), TG (tegument), and AC (acetabulum)]

Fig. 10

Scanning Electron Micrograph (SEM) of the adult P. cervi treated with albendazole (a, b, and c) compared with untreated P. cervi (7a, 7b, and 7c) [Here, AS (anterior sucker), P (papillae), GP (gonopore), TG (tegument), and AC (acetabulum)]