The effects of combination of methanolic leaf extract of Azadirachta indica and diminazene diaceturate in the treatment of experimental Trypanosoma brucei brucei infection in rats (original) (raw)
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Comparative Clinical Pathology, 2012
This study investigated the erythrocytic profile of rats experimentally infected with Trypanosoma brucei brucei and treated with a combination of methanolic leaf extract of Azadirachta indica and diminazene diaceturate (DDA). Acute toxicity study of the drug and extract combinations was carried; selection of the best drug and extract combinations was carried out using 54 rats of both sexes separated into nine groups. Three dose combinations were derived from the selection of the best drug and extract combinations used for the final study, viz: 7 mg/kg body weight (bw) DDA plus 125 mg/kg bw extract (group B), 3.5 mg/kg bw DDA plus 250 mg/kg bw extract (group C) and 1.8 mg/kg bw DDA plus 500 mg/kg bw extract (group D). The final study had, in addition to the three groups derived from the dose-response study, four other groups, viz: uninfected untreated negative control (group F), infected and treated with 3,000 mg/kg bw extract alone (group E), infected and treated with 7 mg/kg bw DDA alone (group A) and infected untreated positive control (group G). The parameters assessed were onset of parasitaemia (OP), level of parasitaemia (LOP), clearance of parasites posttreatment (COPPT), relapse of infection period (RIP), red blood cell counts (RBC) and packed cell volume (PCV). There was no significant difference (p<0.05) in OP between the groups. A day following treatment, the LOP of groups A, B and C was found to be significantly lower (p<0.05) than that of group D (p<0.05) which in turn was lower (p<0.05) than that of group E and G, respectively. The mean LOP of group E was significantly (p<0.05) lower than group G (p<0.05) 2 days posttreatment, and this trend continued throughout the experimental period. Mean COPPT of group D was significantly (p<0.05) longer than that of groups A, C and B. There was no significant difference (p<0.05) in the mean COPPT among groups B, C and A. The mean RIP of group D was significantly shorter (p<0.05) than group C, and that of group C was significantly shorter (p<0.05) than group A. There was no relapse of infection in group B rats. Group B rats had significantly higher (p<0.05) PCV and RBC counts when compared to other infected groups. Group E rats had significantly higher (p<0.05) PCV and RBC counts when compared to group G rats. It was concluded that dose combination of 125 mg/kg bw extract plus 7 mg/kg bw DDA led to significant enhancement of erythrocytic profile and potentiation of diminazene in its trypanocidal activity. This combination therapy proved to be better than single therapy of DDA.
Comparative Clinical Pathology, 2012
This study investigated the leucocytic profile of rats experimentally infected with Trypanosoma brucei brucei and treated with a combination of methanolic Azadirachta indica leaf extracts (MAILE) plus diminazene diaceturate (DDA). Acute toxicity study of the drug and extract combinations was carried. Selection of the best drug and extract combinations was carried out using 54 rats of both sexes separated into nine groups. Three dose combinations were derived from the selection of the best drug and extract combinations used for the final study viz, 7 mg/kg body weight (bw) DDA plus 125 mg/kg bw extract (group B), 3.5 mg/kg bw DDA plus 250 mg/kg bw extract (group C) and 1.8 mg/kg bw DDA plus 500 mg/kg bw extract (group D). The final study had in addition to the three groups derived from the dose-response study, four other groups viz, uninfected untreated negative control (group F), infected and treated with 3,000 mg/kg bw extract alone (group E), infected and treated with 7 mg/kg bw DDA alone (group A) and infected untreated positive control
Journal of Ethnopharmacology, 2010
Aim of the study: To determine the toxicity and anti-trypanosomal activity of the ethanolic extract of Azadirachta indica (Maliacea) stem bark, through in vivo and in vitro approach using Trypanosoma brucei brucei. Materials and methods: Graded concentrations (100, 200, 400, 800, 1600 and 3200 mg/kg) of the crude stem bark ethanolic extract of Azadirachta indica, Hochst ex. A. Dc. (Maliacea) was tested for acute toxicity in 35 out bred Swiss (Wister) adult albino rats of both sexes. Secondly, the in vitro activity in test tubes and in vivo activity of the extract in 30 out bred Swiss (Wister) adult albino rats against Trypanosoma brucei brucei strain NITR/14 (Federe) was evaluated in a graded dose manner. Results: The calculated intra-peritoneal LD 50 of the extract was 870 mg/kg and produced toxicity at high doses (>800 mg/kg). Graded concentrations of the ethanolic extract produced remarkable in vitro activity against Trypanosoma brucei brucei within seconds of inoculation. It also suppressed the establishment of parasitaemia at 100 mg/kg when administered simultaneously with infection in vivo. Similarly, at 200 and 400 mg/kg, the extract administered at the onset of parasitaemia for 4 consecutive days reduced parasitaemia, modulated declined packed volume (PCV) changes by day 48 post-infection in vivo. Conclusion: The results confirm that the folkloric medicinal application of the extract of Azadirachta indica (Maliacea) has a pharmacological basis. Further investigation is however, needed to optimize the effectiveness of the extract.
The effect of Cymbopogoncitratus (lemon grass) extract and berenil was investigated on albino rats, infected with Trypanosoma brucei brucei in the Laboratory. A total of thirty five (35) rats were used, 8-10 weeks old weighing between 120 to 180grams were allowed to acclimatizefor a period of one week. These were divided into seven groups (A and B controls, while C, D, E, F and G groups were infected and treated with 0.5ml and 0.2ml of the aqueous extract of C. citratus and berenil respectively. One milliliter (1ml) of T.b. brucei infected blood was diluted with phosphate buffered saline and was inoculated via the intra-peritoneal route; parasite was detected by wet film method and examined under the light microscopic field using X40 magnification to check thewriggling movement of trypanosomes. Treatment C. had the highest mean PCV (52.93±4.23%) while B. had the least (49.60±4.611%). A. had the highest mean white blood cell value (9.21±1.959x10 9 /L) while D. had the least (8.56±2.915X10 9 /L). Treatment A. also had the highest mean red blood cell value (2.58±1.959X10 9 /L), G. (2.50±3.615X10 9 /L), while C. had the least (2.38±3.869X10 9 /L). Treatment B. had the highest mean weight gain (3.99±0.201g), A. (2.73±0.462g) while E. had the least (0.20±0.346g). Group B. had the highest mean parasitaemia count (77.75±36.130), C. (35.75±40.419) while A. and D. had the least (0.00±0.000). Mortality rate was higher in group B.
Pharmacognosy Research, 2015
Introduction: The medicinal properties of Azadirachta indica have been harnessed for many years in the treatment of many diseases in both humans and animals. Materials and Methods: Twenty-five apparently healthy dogs weighing between 3 and 8 kg were randomly divided into five groups with five dogs in each group. Ameliorative effect of A. indica on erythrocyte antioxidant status and markers of oxidative stress were assessed. Liver and kidney function tests were also performed. Results: Pre-treatment with methanolic extract of Azadirachta indica (MEAI) at different doses did not significantly alter the values of alanine aminotransferase, aspartate aminotransferase and alkaline phosphatase activity in Trypanosoma brucei infection. Although, serum creatinine significantly (P < 0.05) decreased with pre-treatment with 50 mg/kg A. indica, after 2 weeks of T. brucei infection. However, the reduced glutathione (GSH) content of the erythrocyte increased significantly in animals pre-treated with 50 mg/kg and 200 mg/kg of A. indica respectively. Markers of oxidative stress such as malondialdehyde and hydrogen peroxide generated were higher in animals infected with T. brucei with no significant (P >0.05) difference compared to the values obtained in pre-treated animals. Pre-treatment with 100 mg/kg and 200 mg/kg of A. indica significantly (P < 0.05) decreased serum myeloperoxidase activity at 2 weeks post-infection with T. brucei. Conclusion: From this study, MEAI showed significant ability to attenuate oxidative stress and inflammation during experimental T. brucei infection.
Mini Review:Toxicity Study Of Plant Extracts
2020
Treatment of diseasesusing plant-based medicines hadincreased significantly which waspredicted due to its fewer side effects. However, some of the medicinal plantsmight not be scientifically tested, hence their side effects remainunknown. It is necessary to carry out a toxicity study to ensure the safety ofthe plants in the animal models.This review article aims to provide comprehensive information about the toxicity study of medicinal plants. Various factors could influence the results of the assay,starting from the harvesting of the plants,the drying, the plant extraction, the determining of doses used, the animals housing and feeding conditions, the measurement of body and organ weight, the serum preparation and organ isolation, biochemical, hematology, and histopathology parameters.The most influential factor in toxicity study is the dose of the plant extracts.
The comparative efficacy of diaminazine aceturate (DA) and isomethamedium chloride (IC) was investigated in Trypanosoma brucei brucei infected domestic rabbits (Oryctolagus cunisculus). A total of eighteen rabbits were used for the study. The rabbits were divided into six groups of three each. All the rabbits in groups B – F were infected with Trypanosoma brucei brucei, while those in group A served as the negative control (uninfected and untreated group). Group B contained the infected and untreated (positive control group), Group C rabbits were infected and treated with DA at 3.5 mg/kg, Group D were infected and treated with DA 7 mg/kg, Group E contained the infected and treated with 3.5 mg/kg of DA combined with 1.0 mg/kg of IC and Group F contained the infected and treated with 7 mg/kg of DA combined with 1.0 mg/kg of IC. The weekly rectal temperature, body weight gain, packed cell volume (PCV), total erythrocyte count, haemoglobin concentration (HbC), total leucocyte count (TLC), differential leucocyte count (DLC), clinical signs and survivability and rate of parasite clearance were used to assess the efficacy of the drugs and drug combinations. The parasites cleared from groups E and F 24 hours post treatment, while in group D, it was 48 hours post treatment. The rabbits in group C and group B died within 13 – 19 and 50 – 52 days post infection (PI), respectively. Relapse was recorded in all the rabbits treated only wit DA. There was significant (p<0.05) reductions in weight, PCV, erythrocyte count, HbC and TLC. The significant (p<0.05) increase in temperature following infection were reversed by the treatments, this reversal however, was faster and lasted longer in the combined treatment groups (E and especially F) than in the single treatment group. The results of this study suggest that the combined treatments of DA and IC produced better therapeutic effect than DA only.
Nigerian Journal of Basic and Applied Sciences, 2010
The in vitro trypanocidal activity of aqueous extracts of some selected medicinal plants used by local herdsmen in the treatment of various animal diseases in Sokoto State, Nigeria was conducted. Trypanosoma brucei brucei were cultured using 96 well micro titer plate and maintained at 37 0 C. About 20-25 parasites per microscope field were dosed with 1, 2 and 4mg/ml of aqueous extracts of the plants and a control group without extracts. After 5 minutes incubation in Eppendorf tubes maintained at 37 0 C, the parasites survived more than four (4) hours in the absence of extract/Berenil. At 4mg/ml of the extracts 0f Terminalia catappa, Waltheria indica, Cucurbita pepo, Entada abyssinica and Ximenia Americana, complete cessation of motility of T. brucei brucei within 60 minutes was observed. However, at 2mg/ml of Waltheria indica, trypanosome motility after 20 minutes was stopped but Terminalia catappa, and Ximenia americana were found to reduce trypanosome motility at 35 and 55 minutes respectively. Only Waltheria indica reduced trypanosome motility within 25 minutes at 1mg/ml concentration. Berenil, the standard drug, caused cessation of trypanosomal motility within 60 minutes even at 1mg/ml. From the results Waltheria indica was the most effective among the extracts when compared with Berenil, and may be a potential source of compounds with trypanocidal activity.
Ethno-veterinary medicine: screening of Nigerian medicinal plants for trypanocidal properties
Journal of …, 2001
Trypanosoma congolense and T. brucei bloodstream form parasites were propagated axenically in suitable standard media at 34°C. The effects of 33 plant extracts, fractions and pure compounds were evaluated on two clones of T. brucei and drug-sensitive and multi-drug-resistant clones of T. congolense. The cytotoxic activity of the trypanocidal extracts was also evaluated on calf aorta endothelial cells in vitro. Of the extracts tested, 22% killed T. congolense IL 1180 at a concentration of 100 mg/ml while 18% killed 90-100% of T. brucei ILTat 1.4 at the same concentration. However, 6% of the active extracts killed 93% of a dyskinetoplastid form of T. brucei IL Tat 1.1, indicating that the intact kinetoplast is a target of some of the compounds tested. Of the 12 extracts that displayed activity against drug sensitive trypanosomes, 66.7% had trypanocidal activity on a multi-drug-resistant clone, T. congolense IL 3338. The extracts of Eugenia uniflora, Acacia artaxacantha, Terminalia i6orensis, T. superba and Alchornea cordifolia had median lethal concentrations of between 13 and 69 mg/ml on both the drug-sensitive, IL 1180 and multi-drug-resistant clone, IL 3338. The median lethal doses of the active plant extracts on the calf aorta endothelial cells varied between 112 and 13 750 mg/ml while the calculated selective indices ranged between 0.71 and 246.8 indicating bright prospects for the development of some of these extracts as potential trypanocidal agents.