Effect of daflon-500®, a flavonoid compound on chlorpyriphos-induced oxidative changes in the hypophysis and testes in adult male rats.

Alteration of redox status is one of the molecular pathways commonly associated with pesticide toxicity. Antioxidants, including those obtained from plant phenolics, have been shown to mitigate pesticide-induced cellular injury. The present study was aimed at evaluating the effect of daflon-500 ® , a flavonoid compound on sub-chronic chlorpyriphos-evoked changes in antioxidant and biochemical parameters in the hypophysis and testes of adult male rats. Twenty-five male albino rats were randomly divided into 5 groups of 5 animals each. Group I (DW) received distilled water (2 ml/kg); group II (SO) was dosed with soya oil (2 ml/kg); Group III (DAF) received daflon-500 ® at 1000 mg/kg ~ 1/5th of LD50 (≥ 5000 mg/kg); group IV (CP) was administered chlorpyriphos at 7.74 mg/kg ~ 1/10th of LD50 (77.4 mg/kg) while group V (DAF + CP) was previously treated with daflon-500 ® (1000 mg/kg) and then exposed to CP (7.74 mg/kg), 30 min later. Daily oral regimen administration was done for 60 days after which the animals were sacrificed by cervical venesection after light chloroform anesthesia. The hypophysis and testicular tissues were harvested, and their homogenates were analyzed for malondialdehyde, catalase and superoxide dismutase, and acetylcholinesterase levels. A significant increase in the hypophysis and testicular MDA concentrations, coupled with a decrease in the SOD, CAT, and AChE activities were observed in the CP group. The levels of these oxidative and biochemical parameters were alleviated in the group pretreated with Daflon-500 ® . Results of this study demonstrated that pre-treatment with Daflon-500 ® mitigated CP-induced alterations in oxidative and biochemical parameters apparently due to the antioxidant effect of the flavonoid compound.

In vitro anthelmintic activity of Dennettia tripetala G. Baker (Annonaceae) Fruits against Haemonchus contortus.

Helminthosis is one of the greatest causes of parasitic disease and loss in animal productivity. As such, the control of helminth parasites is of critical importance. This study was aimed to investigate the in vitro anthelmintic activity of Dennettia tripetala G. Baker (Annonaceae) fruits against Haemonchus contortus. Using in vitro techniques, the anthelmintic activity of extracts and fractions of D. tripetala G. Baker (Annonaceae) was evaluated for ovicidal (Egg hatch inhibition test) and larvicidal (larval mortality test) activity. Besides, the maximum tolerated dose was determined in adult albino rats administered, 300, 400, and 500 mg/kg body weight of the CME fraction, and observed over a period of 48 h for signs of toxicity and mortality. Phytochemical screening uncovered the occurrence of flavonoids, steroids/triterpenes, cardiac glycosides, saponins, tannins, carbohydrates, and alkaloids in the crude methanol extract (CME), the ethyl acetate fraction (EAF), and butanol fraction. The maximum tolerated dose of the CME of D. tripetala did not produce observable signs of toxicity or death in all the rats given up to 500 mg/kg. The CME and EAF of D. tripetala fruits produced a significant ( p < 0.05 ) reduction in the hatchability of H. contortus eggs in a concentration-dependent manner, while the CME at concentrations between 12.5 and 100 mg/ml completely inhibited the hatching of H. contortus eggs. Similarly, EAF at doses of 25, 50, and 100 mg/ml completely inhibited the hatching of H. contortus eggs. The CME and EAF of D. tripetala fruits produced significant ( p < 0.05 ) larvicidal activity against L3 of H. contortus in a concentration-dependent manner while the CME at concentrations between 6.25 and 100 mg/ml caused larval mortality of H. contortus L3 larvae completely. This study suggests that methanol extract and fractions of D. tripetala fruits possess beneficial anthelmintic (ovicidal and larvicidal) activity against H. contortus, and may be a suitable alternative anthelmintic candidate for the control of nematodes.

Antitrypanosomal properties of Anogeissus leiocarpa extracts and their inhibitory effect on trypanosome alternative oxidase.

Background: African trypanosomiasis is a protozoan disease with huge socio-economic burden to sub-Saharan African exceeding US$4.6 annual loss. To mitigate the incidence of trypanosomal drug resistance, efforts are geared towards discovery of molecules, especially from natural products, with potential to inhibit important molecular target (trypanosome alternative oxidase, TAO) in trypanosomes that are critical to their survival. Method: Crude methanol extract of Anogeissus leiocarpa was subjected to in vitro bioassay-guided antitrypanosomal assay to identify the most active extract with trypanocidal activity. The most active extract was run on a column chromatography yielding five fractions, F1-F5. The fractions were assayed for inhibitory effect on TAO. The most promising TAO inhibitor was subjected to antitrypanosomal evaluation by trypanosome count, drug incubation infectivity test (DIIT) and in vivo studies. Gas chromatography-mass spectrometry (GC-MS) was used to identify and quantify phytochemical constituents of the potential TAO-inhibiting fraction. Results: Ethyl acetate extract (EtOAc) significantly (p<0.05) produced trypanocidal effect and was the most active extract. Of the five fractions, only F4 significantly (p<0.05) inhibited TAO compared to the control. F4 completely immobilised the trypanosomes up to 0.5 µg/µl, yielding an EC50 of 0.024 µg/µl compared to the 0.502 µg/µl of diminazene aceturate positive control group. The DIIT showed that F4 was significantly (p<0.05) potent up to 0.1 µg/µl. F4 significantly (p<0.05) suppressed parasite multiplication in systemic circulation of the treated rats and significantly (p<0.05) maintained high PCV when compared to the 5% DMSO group. Furthermore, F4 significantly (p<0.05) lowered serum concentrations of malondialdehyde. Phytoconstituents identified by the GC-MS include tetradecene; cetene; 3-(benzylthio) acrylic acid, methyl ester; 1-octadecene; 9-heptadecanone; hexadecanoic acid, methyl ester; dibutyl phthalate; eicosene; octadecenoic acid, methyl ester; oleic acid; 2-methyl-Z,Z-3,13-octadecadienol; 1-docosene; 3-phenylthiane, s-oxide; phenol, 3-methyl; phthalic acid, di(2-propylpentyl) ester and 1,4-benzenedicarboxylic acid, bis (2-ethylhexyl) ester. Conclusion: F4 from EtOAc contains six carbohydrates (9.58%), two free fatty acids (6.48%), five fatty acid esters (27.73%), two aromatic compounds (50.63%) and one organosulphide (5.61%). It inhibited TAO and demonstrated antitrypanosomal effects.

Partially Purified Leaf Fractions of Azadirachta indica Inhibit Trypanosome Alternative Oxidase and Exert Antitrypanosomal Effects on Trypanosoma congolense.

INTRODUCTION: Trypanosomiasis is a neglected disease of humans and livestock caused by single-celled flagellated haemo-protozoan parasites belonging to the genus Trypanosoma. PURPOSE: Widespread resistance to trypanocidal drugs creates urgent need for new, more effective drugs with potential to inhibit important trypanosome molecular targets. METHODS: Nine column chromatographic, partially purified leaf fractions of Azadirachta indica (AIF) were subjected to trypanosome alternative oxidase (TAO) inhibition assay using ubiquinol oxidase assay. The potent TAO inhibitors were evaluated for trypanocidal activities against T. congolense in rat model using in vitro, ex vivo, and in vivo assays. Complete cessation or reduction in parasite motility was scored from 0 (no parasite) to 6 (greater than or equal to 6 × 107 trypanosomes/milliliter of blood), and was used to evaluate the efficacy of in vitro treatments. RESULTS: Only AIF1, AIF2, and AIF5 significantly inhibited TAO. AIF1 and AIF5 produced significant, dose-dependent suppression of parasite motility reaching score zero within 1 h with EC50 of 0.005 and 0.004 µg/µL, respectively, while trypanosome-laden blood was still at score six with an EC50 of 44,086 µg/µL. Mice inoculated with the concentrations at scores 0 and 1 (1-2 moribund parasites) at the end of the experiment did not develop parasitaemia. The two fractions significantly (p < 0.05) lowered parasite burden, with the AIF5 exhibiting highest in vivo trypanocidal effects. Packed cell volume was significantly higher in AIF1 (p < 0.05) and AIF5 (p < 0.001) groups compared to DMSO-treated group. Only AIF5 significantly (p < 0.05) lowered malondialdehyde. CONCLUSION: AIF1 and AIF5 offer prospects for the discovery of TAO inhibitor(s).

Anthelmintic activity of methanol extract of Dennettia tripetala G. Baker (Annonaceae) fruits against Haemonchus contortus in red Sokoto goats.

Haemonchosis is a gastrointestinal parasitic disease of economic importance in ruminants especially sheep and goats. In view of the rising costs of conventional veterinary anthelmintics and the development of resistance by Haemonchus contortus, there is a need to develop alternative ethnoveterinary therapies for the treatment and management of Haemonchosis. This study investigated the anthelmintic activity of Dennettia tripetala G. Baker (Annonaceae) fruits against Haemonchus contortus in red Sokoto goats. The maximum tolerated dose was determined in adult albino rats administered, 3000, 4000, and 5000 mg/kg body weight of the crude methanol extract (CME) fraction, and observed over a period of 48 h for signs of toxicity and mortality. The in vivo anthelmintic activity was evaluated using 20 kids infected with H. contortus and randomly allocated into 5 groups (1, 2, 3, 4, and 5). Kids in groups 1, 2, and 3 were treated with CME at doses of 1250, 2500, and 5000 mg/kg respectively for 3 consecutive days per os. Groups 4 and 5 were treated with albendazole (7.5 mg/kg) once and distilled water (5 ml) respectively and served as treated and untreated controls. Blood samples were collected for haematology. The maximum tolerated dose of the CME of D. tripetala did not produce observable signs of toxicity or death in all the rats given up to 5000 mg/kg. There was significant (P < 0.05) reduction in faecal egg count by CME at doses of 1250 mg/kg (91.6%), 2500 mg/kg (98.5%), and 5000 mg/kg (100%) at day 14 post-treatment. The total plasma protein (TPP) and FAMACHA© scoring values were significantly improved (P < 0.05) in the treated groups. These results indicate that methanol extract and fractions of D. tripetala fruits possess beneficial anthelmintic activity against H. contortus and may be a suitable alternative anthelmintic candidate for the control of haemonchosis in goats.

Antiulcer Effects of Methanol Extract of Euphorbia hirta and Honey Combination in Rats.

Stomach ulcer is an endemic gastrointestinal disorder which constitutes a major public health problem all over the world. Stomach ulcer results when there is an imbalance between the protective factors (mucus and bicarbonate) and aggressive factors (acid and pepsin) in the stomach. Dried powdered leaves and stem of the phytomedicine Euphorbia hirta (E. hirta) (1000 g) was extracted with methanol using a soxhlet apparatus. The evaluation of the phytochemical constituents of E. hirta and acute toxicity (to ascertain the safety of using the phytomedicine over a short period of time) was carried out. The antiulcer and gastroprotective effects of crude extract of E. hirta combined with honey in rats were evaluated. The study model using 0.6 M HCl model of ulceration was used to evaluate the antiulcer and gastroprotective activities of the phytomedicine. The soxhlet extraction of E. hirta gave a yield of 54.5 g of crude extract (5.45%). Phytochemical screening of E. hirta showed that the extract contains alkaloids, tannins, saponins, glycosides, flavonoids, and unsaturated steroids. Acute toxicity studies showed that LD50 was greater than 5000 mg/kg. The study showed that the crude extract of E. hirta at 200 mg/kg when administered alone had 54% inhibition of ulceration while when administered together with honey increased to 94% inhibition of ulceration, but honey alone had 89.47% inhibition of ulceration. This implied that E. hirta when combined with honey had a synergistic effect and enhanced the inhibition of ulceration, and this could be seen by the protection of the gastric mucosa. The study of the phytomedicine E. hirta combined with honey revealed that the phytomedicine has antiulcer activities against 0.6 M HCl-induced gastric ulcer in rats. This therefore validates usage and claim by the Igbo people of the southeastern part of Nigeria that the phytomedicine of E. hirta combined with honey has good antiulcer potential.

In vitro and in vivo antitrypanosomal efficacy of combination therapy of Anogeissus leiocarpus, Khaya senegalensis and potash.

ETHNOPHARMACOLOGICAL RELEVANCE: Pastoralists in Nigeria mix barks of Anogeissus leiocarpus (AL) Khaya senegalensis (KS) and potash (Pt) to treat animal African trypanosomosis. AIM: To evaluate antitrypanosomal potential of A. leiocarpus, K. senegalensis and potash for insights into the traditional claim of antitrypanosomal combination therapy (ATCT). MATERIALS AND METHODS: Fifty microliter each of six different concentrations of AL, KS, Pt, AL + KS, AL + KS + Pt and diminazene aceturate (DA, positive control) was incubated with 50 µL of parasite-laden blood containing 108Trypanosoma congolense cells in a 96-well microtitre plate. Negative control wells were devoid of the extracts and drug but supplemented with phosphate-buffered saline (PBS). Efficacy of treatment was observed at 1 h interval for complete immobilisation or reduced motility of the parasites. Each incubated mixture was inoculated into mouse at the point of complete immobilisation of parasite motility or at the end of 6-h observation period for concentrations that did not immobilise the parasites completely. For in vivo assessment, thirty-five parasitaemic rats were randomly allocated into seven groups of 5 rats each. Each rat in groups I-V was treated with 500 mg/kg of AL, KS, Pt, AL + KS and AL + KS + Pt, respectively, for 7 days. Rats in groups VI and VII were treated with diminazene aceturate 3.5 mg/kg once and PBS 2 mL/kg (7 days), which served as positive and negative controls, respectively. Daily monitoring of parasitaemia through the tail vein, packed cell volume and malondialdehyde were used to assess efficacy of the treatments. RESULTS: The AL + KS + Pt group significantly (p < 0.05) and dose-dependently reduced parasite motility and completely immobilized the parasites at 10, 5 and 2.5 µg/µL with an IC50 of 9.1×10-4 µg/µL. All the mice with conditions that produced complete cessation of parasite motility did not develop parasitaemia within one month of observation. The AL + KS group significantly (p < 0.05) lowered the level of parasitaemia and MDA, and significantly (p < 0.05) maintained higher PCV than PBS group. CONCLUSION: The combination of A. leiocarpus and K. senegalensis showed better antitrypanosomal effects than single drug treatment and offers prospects for ATCT. Our findings support ethnopharmacological use of combined barks of A. leiocarpus and K. senegalensis by pastoralist in the treatment of animal African trypanosomosis in Nigeria.

A controlled study to investigate anti-diarrhoeal effect of the stem-bark fractions of Terminalia avicennioides in laboratory animal models.

Due to the shortcomings associated with modern synthetic antidiarrhoeal drugs, it is important to find newer, safer and cheaper antidiarrhoeal agents from natural sources. The study was conducted to evaluate the anti-diarrhoeal activity of the fractions of the stem-bark of Terminalia avicennioides in laboratory animal models. The effect of different concentrations (1.0 × 10-3, 2.0 × 10-3, 4.0 × 10-3 and 8.0 × 10-3 mg/mL) of the aqueous methanol (AMF), ethyl acetate (EAF) and hexane (HXF) fractions of T. avicennioides were tested against spontaneous and acetylcholine-induced contractions of rabbit jejunum as well as on histamine-induced contraction of guinea pig ileum. Similarly, the effects of the AMF on gastro-intestinal transit time, castor oil-induced diarrhoea and castor oil-induced enteropooling were evaluated. The AMF, EAF and HXF at concentrations of 1.0 × 10-3, 2.0 × 10-3, 4.0 × 10-3 and 8.0 × 10-3 mg/mL attenuated the contractile effects of both the spontaneous and acetylcholine-induced contractions of rabbit jejunum and that of histamine-induced contraction of guinea pig ileum in a concentration-dependent manner. The AMF at doses of 200, 300 and 500 mg/kg produced significant (p < 0.05) reductions in gastrointestinal transit time of charcoal and incidence of castor oil-induced diarrhoea in mice relative to the untreated control. Similarly, at doses of 300 and 500 mg/kg, AMF significantly (p < 0.05) reduced the weight and volume of intestinal fluid in the treated mice when compared to the untreated animals. The results of this study showed that the stem-bark of T. avicennioides possesses spasmolytic effect and could be a potential antidiarrhoeal agent. However, detailed pharmacological trials are required to justify the clinical use of the plant for treating diarrhoea.

A controlled study to determine the efficacy of Loxostylis alata (Anacardiaceae) in the treatment of aspergillus in a chicken (Gallus domesticus) model in comparison to ketoconazole.

BACKGROUND: The poultry industry due to intensive methods of farming is burdened with losses from numerous infectious agents, of which one is the fungus Aspergillus fumigatus. In a preliminary study, the extracts of Loxostylis alata A. Spreng, ex Rchb. showed good activity in vitro against A. fumigatus with a minimum inhibitory concentration of 0.07 mg/ml. For this study crude, a crude acetone extract of L. alata leaves was evaluated for its acute toxicity in a healthy chicken model and for efficacy in an infectious model of aspergillosis (A. fumigatus). RESULTS: At a dose of 300 mg/kg, the extract induced some toxicity characterised by decreased feed intake and weight loss. Consequently, 100 and 200 mg/kg were used to ascertain efficacy in the infectious model. The plant extract significantly reduced clinical disease in comparison to the control in a dose dependant manner. The extract was as effective as the positive control ketoconazole dosed at 60 mg/kg. CONCLUSIONS: The results indicate that a crude extract of L. alata leaves has potential as an antifungal agent to protect poultry against avian aspergillosis.