In Vitro Anthelmintic Activity of Ethanol Stem Bark Extract of Albizia ferruginea (Guill. & Perr.) Benth

Abstract

Albizia ferruginea (Guill. & Perr.) Benth bark is used in the traditional medicine as a vermifuge. This study sought to determine the anthelmintic activity of the stem bark extract of Albizia ferruginea. The powdered A. ferruginea stem bark was extracted with ethanol. Phytochemical screening was carried out on A. ferruginea ethanol extract (AFE) and then screened for its anthelmintic property against Pheretima posthuma and Haemonchus contortus using the adult motility assay. The effect of AFE and its fractions on the anthelminthic activity of mebendazole and albendazole were also determined using the adult worm (P. posthuma) motility assay. AFE showed a dose-dependent anthelmintic activity against P. posthuma and H. contortus. The least concentration of AFE (0.5 mg/mL) paralyzed and killed P. posthuma within 272.50 ± 12.42 min and 354.50 ± 5.06 min of exposure, respectively. AFE at the least test concentration (0.14 mg/mL) caused paralysis and induced death of H. contortus, after at 63.50 ± 2.98 and 254.96 ± 2.44 min of exposure, respectively. AFE extract at 0.25 and 0.125 mg/mL increased the paralytic and helminthicidal activities of albendazole. The paralytic and helminthicidal activities of mebendazole were reduced when combined with AFE (0.25 and 0.125 mg/mL). Among the three fractions obtained from AFE, the methanol fraction showed the highest anthelmintic activity. The methanol fraction at 0.5 mg/mL caused paralysis after 69.90 ± 0.15 min and death of worm after 92.53 ± 0.74 min of exposure. The petroleum ether and ethyl acetate fractions showed relatively low anthelmintic activity. Phytochemical screening of AFE revealed the presence of tannins, saponins, glycosides, alkaloids, and coumarins. The results from this study show that A. ferruginea possesses anthelmintic activity which gives credence to its folkloric use.

Figure 1

Effect of AFE and albendazole on paralysis and death of P. posthuma. (a) Paralysis time. (b) Death time. No paralysis and death was observed after the maximum time of exposure (8 hours) in the negative control group (Ringer's lactate solution). AFE: ethanol stem bark extract of A. ferruginea.

Figure 2

Effect of AFE on paralysis and death time of H. contortus. (a) Paralysis time. (b) Death time. ALB: albendazole; n = 9, values are mean ± SEM; ^∗∗∗∗p < 0.0001, compared to the control (one-way ANOVA followed by Dunnett's post hoc test). No paralysis was observed after maximum time of exposure (8 hours) in the negative control group (Ringer's lactate solution).

Figure 3

Effect of AFE on paralytic and helminthicidal activity of albendazole. (a, b) Effect of 0.25 mg/mL AFE on albendazole. (c, d) Effect of 0.125 mg/mL AFE on albendazole. ALB: albendazole; AFE: A. ferruginea ethanol stem bark extract; ^∗∗∗∗p < 0.0001, ^∗∗∗p < 0.001, and ^∗∗p < 0.01 compared to control (one-way ANOVA followed by Dunnett's post hoc test).

Figure 4

Effect of AFE on paralytic and helminthicidal activity of mebendazole. (a, b) Effect of 0.25 mg/mL AFE on mebendazole.(c, d) Effect of 0.125 mg/mL AFE on mebendazole. MEB: mebendazole; AFE: A. ferruginea ethanol stem bark extract; ^∗∗∗∗p < 0.0001, ^∗∗∗p < 0.001 compared to the control (one-way ANOVA followed by Dunnett's post hoc test).

Figure 5

Comparison of the effect of AFE and its fractions (methanol, ethyl acetate, and petroleum ether fractions) on paralysis and death time of P. posthuma. (a, b) Paralysis and death time of methanol fraction, respectively. (c, d) Paralysis and death time of ethyl acetate fraction, respectively. (e, f) Paralysis and death time of petroleum ether fraction, respectively; ^∗∗∗∗p < 0.0001, ^∗∗∗p < 0.001, ^∗∗p < 0.01, and ^∗p < 0.05 compared to control (one-way ANOVA followed by Dunnett's post hoc test). AFE: A. ferruginea ethanol stem bark extract; MEF: methanol fraction of AFE; EAF: ethyl acetate fraction of AFE; PEF: petroleum ether fraction of AFE.