Diethyl ether

Sherman is a foldable metal trap designed to capture live animals. Four trap lines were established at a distance of 30 to 50 m, each with 50 traps separated by 10 m. Sites received six days of trapping with a total of 200 traps. All traps were equipped with bait (peanut butter mixed with maize flour and avocado) and placed in the shade to avoid being too hot. Traps were inspected and re-baited (i.e., if the bait was found eaten by insects) every morning between 07:00 and 08:00 h. Each Sherman trap that captured an animal was replaced with a new one (i.e., previously washed). Each individual trapped rodent was first placed in a cotton bag and humanely anesthetized using diethyl ether (LOBA CHEMIEPVT. LTD.) on cotton wool, then placed in a bucket with a lid. Standard external body measurements (body, tail, hindfoot, and left ear), sex, and weight of the trapped hosts were recorded. Species identification followed Happold and Kingdon [53 ], and sex identification followed Hoffmann et al. [54 ]. The rodent species identification was confirmed using molecular (Cytochrome b) techniques.

Saponin content was determined using the methods of Ejikeme et al. [33 ] and Obadoni and Ochuko [34 (link)]. The process involved mixing 100 mL of 20% aqueous ethanol (Fisher Scientific, UK) and the extracts in a 250 mL conical flask. A water bath (Memmert, Germany) was used to heat the mixture at 55°C for 4 hours. The mixture was filtered and the residue was collected, extracted with ethanol, and rotary evaporated. 20 mL of diethyl ether (LOBA Chemie, India) was mixed with the concentrate in a 250 mL separating funnel. The aqueous layer was collected, and the process was repeated before 60 mL of n-butanol (LOBA Chemie, India) and 10 mL of 5% w/v sodium chloride (Rankem, India) were added. The sodium chloride layer was discarded, and the residual solution was heated in a water bath for half an hour before being transferred into a crucible and dried in an oven. The saponin content was calculated as a percentage using the formula below [37 ]: $\begin{array}{c}\%\text{\hspace{0.17em}Saponin}=\frac{\text{Weight\hspace{0.17em}of\hspace{0.17em}Saponin}}{\text{Weight\hspace{0.17em}of\hspace{0.17em}sample}}\times 100.\end{array}$