Anthelmintic Activity of Ethanolic and Aqueous Extracts of Khaya grandifoliola Stem Bark against Heligmosomoides polygyrus: In Vitro and In Silico Approaches.

Background: Parasitic infection remains a serious health trade for humans and livestock. The purpose of this study was to present scientific proof of the anthelmintic properties of Khaya grandifoliola, which the native population uses to cure helminthiasis. Method: Fresh Heligmosomoides polygyrus eggs were isolated from faecal samples of experimentally infected mice. The faecal material was cultured, and L1 and L2 larval stages were recovered after 48 and 120 hours, respectively. Using the worm microtracker, the anthelminthic efficacy of the extracts against H. polygyrus was assessed. Two different extracts (aqueous and ethanol extracts) were prepared. For the ovicidal and larvicidal activities, 100 µL of various concentrations of plant extracts, levamisole and 1.5% dimethyl sulfoxide (DMSO), were introduced into a 96-well microplate titer followed by the addition of 100 µL of embryonated eggs (60 eggs) for the ovicidal activity and 100 µL of L 1 or L 2 larvae (50 larvae) for the larvicidal activity. The movement of the worm was monitored for 24 hours in the worm microtracker at 27°C. The Glide module of the Schrodinger Maestro software was used to perform docking studies. Results: For the aqueous extracts, the highest percentage of inhibition of hatching was 42.77 ± 12% at 7.5 mg/mL. The IC50 values for the ethanol (0.36 mg/mL) extract showed that the ethanol extract had a good inhibitory effect on the ability of parasites to hatch from eggs. The inhibition percentage of L1 larvae motility at 7.5 mg/mL was 98.0 ± 1.66% and 83.33 ± 1.66% for ethanol and aqueous extracts, respectively. The negative controls, distilled water and 1.5% DMSO, had no inhibitory impact on larvae. On L1-larvae, the drug of choice levamisole (positive control) had the highest percentage effect (100.0%). Six compounds had the highest docking score and their interactions with the receptor as well. Grandiamide A interacts most with tyrosine, glycine, phenylalanine, asparagine, and serine, and its benzene ring and oxygens inhibit these receptors. Carbonyl and hydroxyl (OH) groups connect grandiamide D to asparagine, isoleucine, and phenylalanine, respectively. By donating hydrogen to the receptor through OH groups, D-glucopyranose-6-phosphate also forms relatively strong hydrogen bonds with its oxygen-bound phosphorus and the receptor. 1-O-deacetylkhayanolide E interacts most with serine and glutamic acid. The carbamic acid benzyl ester of carbamic acid [(1S)-1-phenyl-2-[(4-methylphenyl) sulfinyl] ethyl] interacts most with the receptor with carbonyl groups and with asparagine and serine. With its abundant hydroxide, D-mannitol acts as a hydrogen donor and acceptor and interacts most strongly with amino acids such as glycine, asparagine, aspartic acid, alanine, and glutamic acid. Conclusions: K. grandifoliola extracts possess anthelminthic properties. However, in vivo studies are still necessary to demonstrate the effectiveness of this plant for the treatment of helminthiasis.

Antimalarial Efficacy of Ethanol Extract of Bridelia micrantha Stem Bark against Plasmodium berghei-Infected Mice.

Background: The spread of drug resistance is a significant issue, particularly in endemic countries with limited resources. The aim of this study was to evaluate antimalarial and antioxidant activity of B. micrantha in order to justify its use in traditional medicine. Methods: Evaluation of the in vivo antimalarial activity of B. micrantha was carried out according to the model of the suppressive and curative test of Peters' over 4 days in infected Swiss albino mice. Antioxidant parameters and stress were measured after intraperitoneal administration of 1 × 107 infected red blood cells. Results: At doses of 150 mg/kg, 300 mg/kg, and 600 mg/kg, administration of B. micrantha substantially produced suppression of P. berghei infection by 67.75%, 73.46%, and 78.99%, respectively, while 84.64% of the untreated group (1% DMSO) had suppression from chloroquine. The curative test significantly decreased the levels of parasitaemia and death in the treated groups. Furthermore, after B. micrantha extract was given to infected mice, a noteworthy increase in total protein, aspartate aminotransferase (AST), and alanine aminotransferase (ALT) was observed. On the other hand, hepatic catalase (CAT) and superoxide dismutase (SOD) productions were considerably greater than that of the healthy control. Mice had considerably lower levels of nonenzymatic antioxidant markers such as glutathione, NO, and MDA showing that the liver was protected. Conclusion: The infected groups responded favorably to the ethanol extract of B. micrantha. This result justifies investigation for its use in Cameroon.

Nematocidal Activity of Ethanol and Aqueous Extracts of Persea americana Seeds against Heligmosomoides polygyrus using the Worm Microtracker Method.

Background: Infections with gastrointestinal helminths constitute a serious obstacle to the good health of the local population in most African Countries. The aim of this study was to evaluate the anthelminthic activity of Persea americana ethanol and aqueous extracts against Heligmosomoides polygyrus using the worm microtracker. Method: Aqueous and ethanolic extracts of P. americana were prepared. Different concentrations of the extracts were tested against the egg and larvae stages of H. polygyrus using an automated high-throughput method. Briefly, embryonated eggs and larvae of this parasite were obtained after the incubation of fresh eggs at 25°C for 24, 48, and 96 hours for embryonated eggs, L1 and L2 larvae, respectively. One hundred microliters of the plant extracts at various concentrations were put in contact in a 96-well microplate with a suspension of 100 embryonated eggs in a total volume of 200 µL and incubated in a worm microtracker where the motility of the worms was recorded every 30 minutes for the ovicidal activity. The final tested extract concentration was 5, 2.5, 1.25, 0.625, and 0.3125 mg/mL, whereas ringer solution (0.95%) and 1.5% Dimethyl sulfoxide (DMSO) were used as negative controls and levamisole as positive control. The same method was used for larvicidal activities. The anthelmintic activity was determined using the average movement of the worms in the tested product compared with the negative control (1.5% DMSO and ringer solution). Results: The egg hatching rates of H. polygyrus had IC50 of 0.49 mg/mL (95% confidence interval: 71.70-92.03) and 0.22 mg/mL (95% confidence interval: 74.28-86.18) for the ethanol and aqueous extract, respectively. These IC50 indicate that the aqueous extract is more active for the inhibition of hatching at a 95% confidence interval. The aqueous and ethanol extracts presented mean inhibitory hatching rates of 78.33 ± 1.67% and 75.67 ± 1.15% at 5 mg/mL, respectively, with no significant differences. The highest percentage of inhibition of L1 larva was observed at 5 mg/mL with 89 ± 2.3%and 85 ± 2.7% for the ethanol and aqueous extracts, respectively. The lowest percentage of inhibition was observed at 0.3125 mg/mL, with 54.67 ± 3.38% and 49 ± 2.64% for the ethanol and aqueous extract, respectively. No significant differences were observed between the two extracts at 5 mg/mL with an inhibitory percentage of 90.67 ± 3.05% (ethanol) and 89.33 ± 2.08% (aqueous). Conclusion: Extracts of P. americana seeds possess nematocidal activity, however, further in silico and in vivo investigations are necessary to confirm their anthelminthic activity.

Antihelminthic Activity of Lophira Lanceolata on Heligmosomoides polygyrus Using an Automated High-Throughput Method.

Background: There are about 13 parasitic infections that are responsible for significant morbidity and mortality but have not received the attention they deserve; thus, they are now known as "neglected tropical diseases" (NTDs). This study was aimed at evaluating the antihelminthic activities of Lophira lanceolata using an automated high-throughput method. Methods: The antihelminthic activity effect of the extracts against H. polygyrus was determined using an automated high-throughput method. For the egg-hatching test, 100 µL of embryonated egg suspension (60 eggs) was added to 100 µL of various concentrations of extracts, levamisole, and 1.5% DMSO in a 96-well round-bottom microtitre plate. After mixing, the 96-well microplate was placed in WMicroTracker and incubated for 24 h at 25°C; the movements were recorded every 30 minutes. The same procedure was used for the larval motility assays, where 100 µL of L1 or L2 larvae (50 larvae) were put in contact with 100 µL of various concentrations of extracts. Results: The ovicidal activity (hatching) had an IC50 of 1.4 mg/mL for the ethanol extract. The aqueous and ethanol extracts of L. lanceolata showed larvicidal activity on the L1 larvae with IC50 of 1.85 mg/mL and 2.4 mg/mL, respectively, as well as on the L2 larvae with IC50 values of 1.08 mg/mL and 1.02 mg/mL for the aqueous and ethanol extracts, respectively. These results showed that the aqueous extract exhibited a stronger inhibitory power on the hatching rate of parasites than ethanol extracts, while the contrary effect was observed for the larval motility assays. Conclusion: This study provides scientific data on the use of L. lanceolata by the local population for the treatment of helminthiases. However, in vivo and toxicity tests are necessary to assess its activity and safety.

Antimalarial Efficacy and Antioxidant Activity of Lophira lanceolata Stem Bark Ethanol Extract Using Plasmodium berghei Induced-Malaria in Swiss Albino's Mice.

Background: Malaria remains a major public health problem in the tropical and subtropical regions. This study aimed of investigating the antimalarial and antioxidant activities of ethanol extract of Lophira lanceolata stem bark. Methodology. The antimalarial activity was determined using the Peter 4-days' suppressive and Rane's curative tests on Swiss albino: these mice were infected with 1 × 107 parasitized red blood cells. The percentage reduction of parasitemia was related to each test, and the liver homogenate was used to assay malondialdehyde, superoxide dismutase, nitrogen monoxide, catalase, and glutathione for the evaluation of oxidative stress. During the curative test, blood was collected for hematological parameters, alanine aminotransferase and aspartate aminotransferase to evaluate liver function. Result: The ethanol extract of L. lanceolata showed a dose-dependent suppressive activity with the highest suppression of 88.22% at 500 mg/kg. Suppression produced by the extract was not significantly higher than that of the reference drug with 96.1%. Similarly, the extract at doses 125, 250, and 500 mg/kg showed significant decreases (P < 0.05) in a dose-dependent manner during the curative test. The ethanol extract of L. lanceolata caused a reduction of tissue markers, such as hepatic oxidative stress, as it increased the enzymatic activity of antioxidant enzymes. Conclusion: The ethanol extract of L. lanceolata possesses both antimalarial and antioxidant activities. However, further in vivo toxicity tests are required to guarantee their safety.

Antimalarial and Antioxidant Activities of Ethanolic Stem Bark Extract of Terminalia macroptera in Swiss Albino Mice Infected with Plasmodium berghei.

Background: Reduction of oxidative stress during malaria infection is considered as being of great benefit so long as treatment and drug development approaches are concerned. This study had the aim of evaluating the antimalarial and antioxidant activities of the ethanolic extract of Terminalia macroptera in Swiss albino mice infected with the Plasmodium berghei NK65 strain. Methods: In vivo, the antiplasmodial activity of the plant ethanolic extract was tested in a four-day suppressive and curative assay using P. berghei in Swiss albino mice. The extract was administered to the mice at doses of 125, 250, and 500 mg/kg per day. Then, parameters, such as parasite suppression and survival time of the mice, were evaluated. Furthermore, the effect of plant extract on liver damage, oxidative stress indicators, and lipid profile changes in P. berghei-infected mice were studied. Results: Administration of T. macroptera significantly suppressed P. berghei infection by 55.17%, 70.69%, and 71.10% at doses of 125, 250, and 500 mg/kg, respectively, whereas chloroquine had 84.64% suppression relative to the untreated group 1% Dimethyl sulfoxide (1% DMSO) at day 4 (post-infection) in the four-day suppressive test. This suppression activity rate was dose-dependent. The curative test also presented a significant reduction in parasitemia and an extension of the survival time of the treated groups. Treatment of infected parasitized mice with the extract of T. macroptera had a significant (p < 0.05) reduction in parameters, such as total protein, aspartate aminotransferase, and alanine aminotransferase. Infection may also lead to a significant increase in the enzymatic activity of liver catalase and superoxide dismutase compared with the normal control group. The non-enzymatic antioxidant activity in parasitized mice was significantly reduced in malondialdehyde and increased in glutathione and nitric oxide when compared with the normal control group. Conclusions: These findings support the ethnobotanical use of T. macroptera stem bark as an antimalarial remedy coupled with antioxidant activity. However, further in vivo toxicity tests are required to ascertain its safety.

Antiplasmodial, Antioxidant and Cytotoxicity Activity of Ethanol and Aqueous Extracts of Khaya grandifoliola Stem Bark.

Background: Malaria is a serious public health problem, especially in sub-Saharan Africa. The aim of this study was to scientifically provide baseline information on the use of Khaya grandifoliola stem bark as an antimalaria drug by traditional healers. Method: The stem barks of K.grandifoliola were harvested and dried to obtain powder, and fifty grams of the powder were soaked in ethanol and hot distilled water respectively, for the preparation of ethanol and aqueous extracts, then dried in an oven at 40°C for the ethanol extract and 50°C for the aqueous extract. Plasmodium falciparum strains 3D7 sensitive and Dd2 resistant to chloroquine, were used to evaluate in vitro antiplasmodial activity using SYBR Green. The ability of the extracts to prevent oxidative stress was assessed by trapping 2, 2'-diphenyl-1-picrylhydrazyl (DPPH); nitric oxide, hydrogen peroxide and ferric reducing power. The cytotoxicity test of the extracts was carried out on RAW 264.7 cell lines and on erythrocytes. The data obtained were entered in the Excel software, then in Graph pad where the IC50 was calculated and the curves plotted. Results: The fifty percent inhibition (IC50) of the antiplasmodial activity of the chloroquine-resistant strain PfDd2 were 54.27 ± 2.41 µg/mL and 31.19 ± 4.06 µg/mL respectively, for the aqueous and ethanol extracts. As for the Chloroquino-sensitive Pf3D7, IC50 of 53.06 µg/mL was obtained for the aqueous extract and 28.03 ± 1.90 µg/mL for ethanol. The DPPH radical scavenging activity presented IC50 of 104 µg/mL for the aqueous and 2.617 µg/mL for the ethanol extract; for the Nitric oxide (NO) presented an IC50 of 301 ± 21 µg/mL for the aqueous extract 140.7 ± 21 µg/mL for the ethanol; for hydrogen peroxide the ethanol and aqueous presented IC50 of 845.1 ± 21 µg/mL and 509.4 ± 21 µg/mL respectively. The cytotoxicity on RAW 264.7 cells presented High CC50 in particular >1000 µg/mL and 467.4 µg/mL respectively for the aqueous and ethanol extract. Conclusion: Extracts of Khaya grandifoliola exhibited antiplasmodial activity. The ability to inhibit oxidative stress as well as lower cell toxicity on RAW 264.7 and erythrocytes, is a good indicator. However, in vivo tests remain important in order to confirm the use of this plant for the treatment of malaria.

Efficacy of Khaya grandifoliola Stem Bark Ethanol Extract in the Treatment of Cerebral Malaria in Swiss albino Mice Using Plasmodium berghei NK65 Strain.

Background: Cerebral malaria is one of the most severe and dangerous forms of malaria and is potentially fatal. This study was aimed at evaluating the anticerebral malaria efficacy of Khaya grandifoliola used by traditional healers. Method: Fifty grams of Khaya grandifoliola stem bark was macerated in 1 L ethanol (95%) for 72 h. The filtrate was dried at 40°C until the obtention of a dry extract. The antimalarial test was evaluated using the Peter 4-day suppressive test and the Rane curative test. Mice were group into 6 groups of 6 mice each. For the antioxidant test, parameters such as malondialdehyde (MDA), superoxide dismutase (SOD), glutathione (GSH), catalase (CAT), and nitric oxide (NO) were assessed. The livers of mice were crushed and centrifuged in order to be measured. Aspartate aminotransferase (ASAT) and alanine aminotransferase (ALAT) using the Dutch Diagnostics Kit and blood were collected for haematological parameters. Results: The ethanol extract showed a suppressive activity of 78.12%, 75.30%, and 68.69% at 500 mg/kg, 250 mg/kg, and 125 mg/kg, respectively. Similarly, the curative activity showed a statistically significant reduction in parasitemia (p < 0.05). Antioxidant parameter assays showed a low value of MDA and a high value of SOD, CAT, NO, and GSH in the negative control group. A statistically significant higher values of ASAT and ALAT were observed in the negative control compared to the other test groups (p < 0.05). Haematological parameters showed a statistically significant decrease in white blood cells, red blood cells, haemoglobin, and platelets in the negative control group (p < 0.05). Conclusion: The results of this study justify the traditional usage of Khaya grandifoliola in the treatment of cerebral malaria. However, in vivo toxicity assessment is still necessary to verify its safeness.

Antiplasmodial, Antioxidant, and Cytotoxic Activity of Bridelia micrantha a Cameroonian Medicinal Plant Used for the Treatment of Malaria.

Introduction: Resistance to common antimalarial drugs and persistence of the endemicity of malaria constitute a major public health problem in Cameroon. The aim of this study was to evaluate the in vitro antiplasmodial, antioxidant, and cytotoxic activities of aqueous and ethanol extracts of Bridelia micrantha used by Cameroonian traditional healers for the treatment of malaria. Methods: Aqueous and ethanolic stem bark extracts were prepared according to standard procedures. The SYBR Green method was used for antiplasmodial activity on strains of Plasmodium falciparum sensitive to chloroquine (3D7) and resistant (Dd2). In vitro antioxidant activities of B. micrantha were determined using the scavenging activity of 2,2'-diphenyl-1-picrylhydrazyl, nitric oxide, ferric reducing power, and hydrogen peroxide as well as their cytotoxicity on RAW 264.7 macrophage cells and red blood cells (RBC). Results: The aqueous and ethanol extracts of Bridelia micrantha showed antiplasmodial activity on the 3D7 strain with IC50 of 31.65 ± 0.79 µg/ml and 19.41 ± 2.93 µg/ml, respectively, as well as 37.64 ± 0.77 µg/ml and 36.22 ± 1.04 µg/ml for the Dd2 strain, respectively. The aqueous and ethanol extracts showed free radical scavenging properties. The IC50 aqueous and ethanol extract was approximately 0.0001737 µg/ml, 42.92 µg/ml, 1197 µg/ml, 63.78 µg/ml and 4.617 µg/ml, 429.9 µg/ml, 511 µg/ml, and 69.32 µg/ml for DPPH, NO, H2O2, and FRAP, respectively, which were compared to ascorbic acid (8.610e - 005 µg/ml, 2901 µg/ml, 3237 µg/ml, and 18.57 µg/ml). The aqueous and ethanol extracts of B. micrantha were found to be nontoxic with CC50 values of 950 ± 6.6 µg/ml and 308.3 ± 45.4 µg/ml, respectively. Haemolysis test showed that the two extracts were not toxic. Conclusion: These results suggest that B. micrantha can serve as an antimalarial agent. However, further studies are needed to validate the use of B. micrantha as an antimalarial.

Antimalarial Activity of Ethyl Acetate Extract and Fraction of Bidens pilosa against Plasmodium berghei (ANKA).

BACKGROUND: Malaria is one of the most critical diseases causing about 219 million cases worldwide in developing countries. The spread and development of resistance against chemical antimalarial drugs is one of the major problems associated with malaria control. The present study was to investigate the antimalarial efficacy of ethyl acetate extract and one fraction of Bidens pilosa in vivo in order to support the usage of this plant by traditional healers to treat malaria. METHODS: The extracts were prepared by maceration of B. pilosa leaf powder in ethyl acetate. The liquid filtrate of the extract and the best in vitro antiplasmodial fraction using HPLC were concentrated and evaporated using a rotavapor under vacuum to dryness. The antimalarial activity of B. pilosa plant products were evaluated in vivo against Plasmodium berghei infected mice according to the Peter and Rane test. The antimalarial efficacy of the a selected crude extract (ethyl acetate extract) was evaluated at 125, 250, and 500 mg/kg, while a selected fraction from ethyl acetate extract (fraction 12) was evaluated at 62.5 and 125 mg/kg. Blood from experimental animals was collected to assess hematological parameters. RESULTS: The crude extract of ethyl acetate and fraction 12 demonstrated 100% in vivo parasite suppressive activity at doses of 500 mg/kg and 125 mg/kg, respectively, for the crude extract and fraction 12. The mice treated with 250 and 500 mg/kg had their parasitemia (intraerythrocytic phase of P. Berghei) drop considerably, disappearing by the 8th day in mice receiving 500 mg/kg. The ethyl acetate extract of B. pilosa, fraction 12 showed an even higher antiplasmodial activity. By the 5th day of the experiment, the treatment led to a modification of hematological parameters in mice. The chloroquine (5 mg/kg), fraction 12 (125 mg/kg), and the crude extract (500 mg/kg) groups all survived the 30 days of the experiment, while the negative control group registered 100% of the deaths. CONCLUSION: This study scientifically supports the use of Bidens pilosa leaves in the traditional treatment of malaria. However, the mode of action and in vivo toxicity of the plant still need to be assessed.