A significant elevation in mean inhibition of adult worm motility beginning 2 h post-exposure with 150 mg/ml of V. vinifera
leaf extract and Albendazole. After 4 h of the exposure time, Albendazole and concentrations of 125 and 150 mg/ml of the extract resulted in significantly higher inhibited motility compared to the remaining concentrations below 125 mg/ml. The lower concentration (25 mg/ml) was significantly more lethal than the negative control (RPMI-1640 medium) at 6 h of exposure. Within 6 h after exposure, concentrations of 100, 125 and 150 mg/ml of V. vinifera
leaf extract were more effective against adult worms than the remaining concentrations below 100 mg/ml. As the concentration and duration of exposure increased, the mortality of adult worms also increased causing death for all parasites (P<0.05) (Fig 1, Table 1).
Table 1: In vitro worm lethal effect of Vitis vinifiera leaf extract on D. dendriticum.
Fig 1: (a) D. dendriticum in the liver (arrows). (b) worms with RPMI 1640 medium. (c) worms treated with V. vinifera leaf extract (150 mg/ml) after 6 hr. (d) worms treated with Albendazole after 6 hr.
The problem of resistance to anthelmintic drugs, their toxicity and growing concern about the presence of drug residues in animal products has led to a renewed interest in the use of herbal medicines. The in vitro
tests using free-living stages of parasitic nematodes offer a means of evaluating the anthelmintic activity of new plant compounds (Asase et al., 2005). V. vinifera
leaf extract showed good in vitro
on D. dendriticum
lethal effect. As the concentration and duration of exposure increased. Within 6 h after exposure, a concentration of 100, 125 and 150 mg/ml of V. vinifera
leaf extract was more effective against adult worms than the remaining concentrations below 100 mg/ml (p<0.05). The present result is comparable to those obtained utilizing different kinds of parasites reported by some researchers. Gholami et al., (2012)
concluded that in vitro
the methanolic extract of Vitis vinifera was tested as anti-leech Limnatis nilotica
and found the methanol extract of grape could be presented as a complementary treatment against leech L. nilotica
in future. Waghorn et al., (2006)
reported that the methanolic extract of the grape seed extract is used to treat Ostertagia
and Trichostrongylus colubriformis
in sheep. Abbas et al., (2020)
reported the V. venifera
seeds extract are showed an inhibitory effect on sporulation and damage of Eimeria
oocysts in chickens, as the morphology of oocysts in terms of shape, size and number of sporocysts, so used anticoccidial. It was also mentioned that the aqueous extract of V. vinifera
leaves shows antibacterial activity against Escherichia coli, Enterococcus feacalis, Staphylococcus aureus (Mansour et al., 2011).
Histological preparations showed changes in D. dendriticum
after in vitro
exposure to V. vinifiera
leaf extract at a concentration of 150 mg/ml, RPMI-1640 medium and Albendazole. Histological observations of D. dendriticum
at RPMI-1640 medium, the tegument was seen covering the outer surface of the body of the worm that the tegument layer ridges were intact and thick. The muscular layer was intact. The nucleus and most of the cytoplasm reside in a massive, bulging cell body (Fig 2).
Fig 2: Transverse section of D. dendriticum at normal RPMI-1640 showing the tegument layer (TG), ridges of the tegument (R) and muscular layer (ML). Scale bar = 20µm.
while, in histological observations of D. dendriticum
at a concentration of 150 mg/ml (Fig 3), there was a similarity with Albendazole (Fig 4).
Fig 3: Transverse section of D. dendriticum at 150 mg/ml of V. vinifera leaf extract showing the detachment of ridges of the tegument (arrows) and the muscular layer stringy and wrinkled (asterisk). Scale bar = 20µm.
Fig 4: Transverse section of D. dendriticum at Albendazole showing the detachment of ridges of the tegument (arrows), the muscular layer stringy and wrinkled (asterisk) and blebbing of the muscular layer (circle). Scale bar = 20µm.
The disruption of the apical tegumental layer was eroded so that it looked thinner than the tegumental layer of D. dendriticum
at RPMI-1640 medium and blebbing of the tegument layer. The muscular layer looks stringy and wrinkled. These situations made the muscular layer of D. dendriticum
at a concentration of 150 mg/ml looked shorter than the negative control. Histological observations of D. dendriticum
at a concentration of 150 mg/ml and Albendazole caused damage to the integumentary structure of D. dendriticum
, especially in the tegument layer and muscular layer. The increase in the level of concentration is directly proportional to the increase in damage to the tegument layer and muscular layer compared to the RPMI-1640 medium these results are similar to Becker et al., (1980). Waghorn et al., (2006)
reported that all the grape seed extracts have in vivo
anthelmintic potential on Ostertagia
and Trichostrongylus colubriformis
in sheep. Mansour et al., (2011)
found that the ethanolic and the aqueous extracts of Vitis vinifera
promoted the destruction of cytoplasmic and nuclear membranes of Leishmania infantum
promastigotes and altered the overall shape of the cell. The bioactive compounds responsible for these activities could be antioxidant compounds including flavonoids, anthocyanins, catechin and epicatechins (Kara et al., 2016).
However further studies and in vivo
trials are needed to understand its anthelmintic effect in sheep.