Ethnoveterinary survey of trypanocidal medicinal plants of the beninese pharmacopoeia in the management of bovine trypanosomosis in North Benin (West Africa).

Cattle breeding is of great socio-economic importance for Benin's cattle farmers in general and those of North Benin in particular. The objective of this study is to inventory the natural products of medicinal plants of the Beninese pharmacopoeia for the management of trypanosomes in cattle in North Benin. The methodology consisted of individual and semi-structured interviews with cattle farmers on the use of medicinal plants. A total of 360 cattle farmers were selected and interviewed in twelve villages in four municipalities (Tchaourou, N'dali, Bembèrèkè and Gogounou) in northern Benin. Different quantitative ethnobotanical indices were calculated to determine the level of use of plant species. The Relative Frequency of Citation (RFC), the Informant Consensus Factor (ICF = 0.918) and the Generic Coefficient (Rg = 1.04) were evaluated. The knowledge of medicinal plants was influenced by the level of education and the main activity of those who practiced animal husbandry. The results yielded 48 medicinal plants for veterinary use belonging to 46 genera and 28 families. The Leguminosae family (12.50%) was the most represented. The most cited plants with a RFC above 10% were K. senegalensis, P. africana, K. africana, M. inermis, S. latifolius, M. polyandra. The parts used were leaves (46.15%); barks (24.62%) and roots (15.38%). Decoction (53.23%), plundering (32.26%) and maceration (11.26%) were the main methods of preparation. The administration was mainly by oral route. The calculated indices show a high diversity of medicinal plants with trypanocidal properties in the control of cattle trypanosomosis in the Sudanese and Sudano-Guinean zones of northern Benin. Plant species with high citation and RFC values should be selected for comprehensive pharmacological and phytochemical research to validate this ethnomedical knowledge in the management of cattle trypanosomosis.

UPLC-QToF-ESI-MS identification and anthelmintic activity of Mitragyna inermis (Willd.) Kuntze (Rubiaceae).

Medicinal plants attract the attention of many researchers to find natural and safe remedies for various resistant diseases. Leaves of Mitragyna inermis are widely used in traditional veterinary medicine for the treatment of gastrointestinal strongyles of small ruminants. The aim of the current study is to estimate the antioxidant, anthelmintic and the larval toxicity of the aqueous and hydroethanolic extracts of this plant in addition to the hexane, dichloromethane and ethanol fractions of the hydroethanolic extract. Investigation of the most active extract using Ultra Performance Liquid Chromatography coupled with Quadrupole Time-of-Flight Electrospray Ionization Mass Spectrometry (UPLC-QToF-ESI-MS). Both plant extracts showed good antioxidant activity by scavenging the 2,2'-diphényl-1-picrylhydrazyl (DPPH) radical and reducing the ferric ion. Similarly, they were no-toxic to Artemia salina larvae (CL50 > 0.1 µg/mL). Also, they significantly reduced larval migration and motility of Haemonchus contortus adult worms (p < 0.001). The hexane, dichloromethane and ethanolic fractions of the hydroethanolic extract showed low activity compared to crude extracts except for the hexane fraction on H. contortus adult worms (p < 0.001) while it showed a poor result on larvae. It thus appears that the anthelmintic activity of the extract may be linked to the synergistic action of these compounds. The UPLC-QToF-ESI-MS analysis revealed the tentative identification of 15 compounds including 7 alkaloids. The results of the present study confirm the anthelmintic activity of M. inermis in traditional veterinary medicine.

Typology of rabbit rearing systems in tropical areas: A case of Benin (West Africa).

The breeding of rabbits in tropical areas is confronted with enormous difficulties related to feeding and health, which limit the expansion and viability of these farms. The objective of this study is to carry out a typology of rabbit farms in tropical conditions to characterize the structure and function of this rearing in order to better understand their production results. A sample of 600 rabbit farms located throughout Benin was selected. The typology was carried out using multiple correspondence analysis (MCA) followed by Hierarchical Cluster Analysis (HCA) according to the Ward's aggregation algorithm and Euclidean distance, which identified five typological groups. Group 1 covered 45.7% of the farms and included small-scale production (fewer than 20 does) by professional breeders using traditional parasite control methods. Group 2 covered 3.3% of the rearing and includes a larger number of semi-extensive farms using self-produced feed. Group 3 (14.7%) was characterized by farms with fewer than 20 does that were semi-extensive and used more phytotherapy. In Group 4 (9.7% of the farms), the extensive mode was the most used, with veterinary medicine being the most used. Group 5 concentrated 26.7% of the farms and was marked by semi-extensive breeding. These farms did not record any cases of parasitosis. The typology carried out allowed a better understanding of the operating modes of these farms, their problems and the main limiting factors.

Anthelmintic Activity, Cytotoxicity, and Phytochemical Screening of Plants Used to Treat Digestive Parasitosis of Small Ruminants in Benin (West Africa).

Medicinal plants continue to be used alone or in combination with veterinary drugs to treat animal ailments, especially in developing countries where livestock farmers often lack access to modern veterinary services and drugs. In addition, digestive parasitosis remain a major constraint for small ruminant livestock. The objective of this study was to screen the anthelmintic activity of the main plants used in the treatment of the digestive parasitosis of small ruminants in Benin. A total of 40 extracts were prepared using the successive maceration of 10 plants in four solvents of increasing polarity. The phytochemical screening of the plants was performed, and the anthelmintic activity of the extracts was evaluated on L3 larvae of Haemonchus contortus. The cytotoxicity of the 40 extracts was determined on WI38 noncancerous fibroblast cells using the MTT assay, and the total phenol content (TPC), total flavonoid content (TFC), and condensed tannin content (CTC) were quantified in the most effective extracts using colorimetric methods. The results show that the plants contained tannins, flavonoids, and triterpenoids which may, in part, justify their anthelmintic activities. All plants gave active extracts at the highest concentration tested (1200 µg/mL). Methanol (MeOH) extracts were, in general, more effective than the hexane (HEX), dichloromethane (DCM), and aqueous (H2O) ones in inhibiting larval migration, with the MeOH extracts of Terminalia leiocarpa, Adansonia digitata, and Momordica charantia being the most effective. Nevertheless, the MeOH extract of M. charantia was highly cytotoxic at the concentration of 100 µg/mL. The anthelmintic activity of M. charantia, Vitex doniana, and Caesalpinia bonduc was studied on H. contortus for the first time. These results provide scientific information that can be used for better valorization of the anthelmintic potential of the studied plants and to initiate the process of the identification of new anthelmintic molecules.

Fractionation Coupled to Molecular Networking: Towards Identification of Anthelmintic Molecules in Terminalia leiocarpa (DC.) Baill.

Terminalia leiocarpa is a medicinal plant widely used in ethnoveterinary medicine to treat digestive parasitosis whose extracts were shown to be active against gastrointestinal nematodes of domestic ruminants. The objective of our study was to identify compounds responsible for this activity. Column fractionation was performed, and the activity of the fractions was assessed in vitro on Haemonchus contortus and Caenorhabditis elegans as well as their cytotoxicity on WI38 fibroblasts. Two fractions were the most active on both nematode models and less cytotoxic. LC-MS/MS analysis and manual dereplication coupled to molecular networking allowed identification of the main compounds: ellagic acid and derivatives, gallic acid, astragalin, rutin, quinic acid, and fructose. Other potentially identified compounds such as shikimic acid, 2,3-(S)-hexahydroxydiphenoyl-D-glucose or an isomer, quercetin-3-O-(6-O-galloyl)-ß-D-galactopyranoside or an isomer, and a trihydroxylated triterpenoid bearing a sugar as rosamultin are reported in this plant for the first time. Evaluation of the anthelmintic activity of the available major compounds showed that ellagic and gallic acids were the most effective in inhibiting the viability of C. elegans. Their quantification in fractions 8 and 9 indicated the presence of about 8.6 and 7.1 µg/mg ellagic acid and about 9.6 and 2.0 µg/mg gallic acid respectively. These concentrations are not sufficient to justify the activity observed. Ellagic acid derivatives and other compounds that were found to be positively correlated with the anthelmintic activity of the fractions may have additive or synergistic effects when combined, but other unidentified compounds could also be implicated in the observed activity.

Anthelmintic flavonoids and other compounds from Combretum glutinosum Perr. ex DC (Combretaceae) leaves.

A chemical study of the hydro-ethanol extract of the leaves of Combretum glutinosum resulted in the isolation of nine compounds, including 5-demethylsinensetin (1), umuhengerin (2), (20S,24R)-ocotillone (3), lupeol (4), ß-sitosterol (5), oleanolic acid (6), betulinic acid (7), corymbosin (8) and ß-sitosterol glucoside (9). Four compounds have been isolated for the first time from the genus Combretum [viz. (1), (2), (3) and (8)]. The crystal structures of flavonoid (2), C20H20O8, Z' = 2, and triterpene (3), C30H50O3, Z' = 1, have been determined for the first time; the latter confirmed the absolute configuration of native (20S,24R)-ocotillone previously derived from the crystal structures of related derivatives. The molecules of (3) are linked into supramolecular chains by intermolecular O-H...O hydrogen bonds. The crude extracts obtained by aqueous decoction and hydro-ethanolic maceration, as well as the nine isolated compounds, were tested for their anthelmintic activity on the larvae and adult worms of Haemonchus contortus, a hematophage that causes parasitic disorders in small ruminants. The evaluated anthelmintic activity showed that the extracts at different doses, as well as all the compounds tested at 150 µg ml-1, inhibited the migration of the larvae and the motility of the adult worms of the parasite compared with the phosphate buffer solution negative reference control. The best activity was obtained with flavonoids (1), (2) and (8) on both stages of the parasite. The flavones that showed good activity can be used for the further development of other derivatives, which could increase the anthelmintic efficacy.

Ethnoveterinary knowledge of sheep and goat farmers in Benin (West Africa): effect of socioeconomic and environmental factors.

Sheep and goats are two of the main animal species raised in Benin and one of the main sources of income for people living in rural areas. Faced with the inaccessibility of synthetic veterinary products and their low purchasing power, the majority of breeders use ethnoveterinary practices to treat small ruminants diseases. The specific objectives of the current study were (1) to document the traditional knowledge regarding the disorder groups treated and the medicinal plants used in the health and zootechnical management of small ruminants in Benin and (2) to assess the effect of gender, ethnicity, agro-ecological zone and herd size associated with them. To achieve these objectives, an ethnoveterinary survey was conducted in different agro-ecological zones from September 2018 to February 2019. A questionnaire was administered to 506 breeders. The data were analyzed through calculation of the Fidelity Level (FL), Cultural Importance Index (CI) and Informant Consensus Factor (ICF). Ten disorder groups were treated by the people surveyed. These were mainly digestive disorders (D) and reproductive disorders (W), both presenting a ICF value of 0.8. A total of 101 medicinal plants belonging to 42 families and 90 genera were recorded. Spondias mombin, Zanthoxylum zanthoxyloides and Khaya senegalensis were the most important plants with CI values of 0.208, 0.125 and 0.121, respectively. Gender, ethnicity, agro-ecological zone and flocks size were the socioeconomic and environmental factors that significantly influenced the level of ethnoveterinary knowledge. Chemical and biological analysis are needed on less studied plants such as, Striga hermonthica, Crossopteryx febrifuga, Elaeis guineensis and Momordica charantia.

In vitro effects of tannin and extracts of Bridelia ferruginea and Mitragyna inermis on the exsheathment of infective larvae of Haemonchuscontortus.

Bridelia ferruginea (Euphorbiaceae) and Mitragyna inermis (Rubiaceae) are two plants of the beninese pharmacopeia used in vivo for the control of gastrointestinal nematodes (GINs) in small ruminants. The objective of the present study is to explore the mechanism of bioactive compounds involved in the action of these two plants on the third-stage infective larvae (L3s) of Haemonchus contortus. Thus, sheathed L3s of H. contortus were incubated with acetone extracts of B. ferruginea and M. inermis at concentrations of 0, 150, 300, 600 and 1200 µg/mL for 3 h at 25°C. The L3s were then washed and artificially submitted to exsheathment in the presence of sodium hypochlorite solution. The role of tannins was verified by adding a tannin inhibitor, polyvinyl polypyrrolidone (PVPP), to the acetone extracts of these two plants for 2 h at 25°C. Acetone extracts from B. ferruginea and M. inermis inhibited the exsheathment of H. contortus larvae (p < 0.001) and this inhibitory effect was dose-dependent for M. inermis at the concentrations tested. Treatment of B. ferruginea and M. inermis extracts with PVPP was associated with a partial restoration of the exsheathment kinetics of H. contortus larvae (p < 0.001), confirming the predominant role of tannins but also the residual role of other secondary metabolites. These in vitro results suggest that these plants are endowed with anthelmintic (AHs) properties and therefore likely to be used as alternatives to synthetic molecules.

Evaluation of the in vivo anthelmintic properties of Mitragyna inermis (Willd.) as a livestock dewormer against parasitic hematophagous worm Haemonchus contortus infections in different breeds of lambs.

Gastrointestinal (GI) nematodes remain a major constraint on livestock production throughout the world. This study assessed the in vivo efficacy of the powder of Mitragyna inermis leaves in three breeds of lambs, namely, West African dwarf lambs (WAD), West African long-legged lambs (WALL), and F1 lambs (cross of a WALL ram with a WAD ewe), artificially infected with 3000 L3s of Haemonchus contortus in a controlled experiment. Fecal sample examination, serological analysis, and necropsy were carried out to determine the egg count, worm burden, and worm fecundity reduction. A dose of 3.2 g/kg body weight (BW) M. inermis was administered per the oral route for three consecutive days and repeated 2 weeks later. Compared with the control, the powder of M. inermis leaves (> 60%) and albendazole (100%) significantly reduced (p < 0.01) fecal egg counts (FECs) in the three breeds of lambs. The posttreatment reduction in FECs fluctuated from 56.99 to 78.75% for WAD lambs, 38.39 to 66.39% for WALL lambs, and 35.55 to 63.11% for F1 lambs (WALL × WAD). Significant differences (p < 0.05) were observed in packed cell volume values before and after infection. M. inermis reduced the egg-laying capacity of female adult worms by up to 60% and eliminated more than 80% of the adult worms of H. contortus in lambs. Furthermore, albendazole reduced the worm count and fecundity of female worms by greater amounts than M. inermis (100%). The findings of this study showed that M. inermis is a good source of bioactive compounds for drug development. According to this result, a 3.2 g/kg BW dose of the plant could be applied for the control of GI nematodes in small ruminants.