Hoof

The geographic coordinates of all houses and vector breeding places were recorded by using a handheld global positioning system receiver with an accuracy of ± 5 m. Census was done at the beginning, on Week 50, and at the end. In the first census, 7,038 (1,212 households) individuals were registered, and the second census added 1,083 subjects—making the total number of studied subjects 8,121 in 1,388 households. The average number of persons per household was 5.9. Figure 2 shows the study profile. The month in which the study subjects came into or left the study area was recorded. The month in which the study subjects left or moved to the study area was considered to contribute 2 weeks to the total number of weeks observed.
Blood samples were collected from all febrile cases using World Health Organization (WHO) standard procedures27 ; a single finger prick was used to take a blood sample for the RDT and to prepare thick and thin blood films for microscopic evaluation. The result of RDT was used to treat the febrile case on the spot. The confirmations of at least two of the three experienced readers were sought to label a study subject as a malaria case. This work dealt with microscopically confirmed (by at least two readers) malaria cases only.
Three major malaria-related interventions (IRS with DDT and later with Deltamethrin and mass ITNs distribution) were carried out by the government within the study period. The brand of the ITNs was PermaNet2.0 (Vestergaard, Frandsen, Switzerland). We did post-intervention surveys to record ITNs and IRS coverage, and document replastering of the insecticide sprayed surfaces in the houses. The efficacies of DDT and Deltamethrin were assumed to be 10% and 50% (taken from Massebo and colleagues, unpublished data of the same study area) on the sprayed week and to decay over time at a constant rate at each time step—becoming nil after 24 weeks and 18 weeks,21 (link),28 respectively. The initial efficacy was multiplied by the coverage minus the proportion of households that practiced replastering of the sprayed surface.
Starting from Week 5, we recorded each household member who slept under the ITNs the night before the interview.
The main vector breeding place was identified by the research team. We explored potential places within and surrounding the Kebele and the place where we found larvae of Anopheles species was the swampy area near the Lake Abaya. There were several small water bodies created mainly by hoof-prints of cattle and hippopotamus. Being on the shore of the lake, these tend to dry slowly after the rainy season—producing extended effect of rainfall. Overflow of the lake during the rainy season followed by shrinkage resulted in more favorable condition for vector breeding.
The meteorological data were obtained from the nearest local meteorological station at Arba Minch University, which was 6 km away from the study area.

Field studies were carried out in Lwanda and Kigoche villages of Homa Bay and Kisumu counties of western Kenya, respectively. Lwanda village is located on the southern shore of the Winam Gulf of Lake Victoria (00°28′28″S, 34°17′22″E) at an altitude of 1,169 m above sea level (Verhulst et al., 2011a (link)). Average rainfall and relative humidity are 1,200 mm and 65%, respectively. The mean temperatures vary between 18°C and 34°C. Hoof prints of cattle and night-grazing hippopotami provide excellent mosquito breeding sites in Lwanda. Fishing and livestock keeping are the main occupation of the local inhabitants. Kigoche village lies adjacent to the Ahero rice irrigation scheme (00°08′19″S, 34°55′50″E) at an altitude of 1,160 m above sea level. Kigoche has an average annual rainfall of 1,000–1,800 mm and an average relative humidity of 65%. Mean annual temperatures in the area vary between 17°C and 32°C. Rice cultivation is the main occupation of the inhabitants. Most houses in the two villages are mud-walled with open eaves, have corrugated iron-sheet roofs, have no ceiling, and are either single- or double-roomed. Eaves, about one foot wide, increase ventilation in the houses and form the predominant entry points for mosquitoes (Snow, 1987 (link); Lindsay and Snow, 1988 (link)). Malaria caused by Plasmodium falciparum is endemic in the two villages. The villages experience two rainy seasons: between April–June and September–October. During these periods, mosquito breeding grounds proliferate, and mosquito populations rapidly increase in size. Cattle, goats, chicken, dogs, cats, and a few sheep constitute the domestic animal population, with cattle being most abundant. Maize, millet and sorghum are cultivated at subsistence level in Lwanda, whereas rice is a main cash crop in Kigoche.
In both villages, trapped mosquitoes were morphologically identified using the keys published by Gillies and Coetzee (1987 ), counted, and the data entered in MS Excel spreadsheets. Culicine mosquitoes were identified up to genus level, and anophelines into An. gambiae sensu lato, An. funestus and other anopheline species. Abdominal statuses of female mosquitoes were categorized as unfed, blood-fed, or gravid. Mosquitoes belonging to the An. gambiae complex were identified to species using a ribosomal DNA Polymerase Chain Reaction assay (Scott et al., 1993 ).

Post-processing of motion capture data was performed using Vicon Nexus software (version 2.10, Vicon Motion System Ltd., Oxford, UK) with 5 trials in which the animal maintained a consistent walking pace reconstructed for each session (8-, 5-, and 1-days pre-stroke and 3 days post-stroke). Markers were labelled and spline and cyclic algorithms used to fill all visible gaps (Vicon Motion System Ltd., Oxford, UK). A fourth order, zero lag, low pass Butterworth filter was applied with a cut-off frequency of 10 Hz. Data were exported to C3D format and further processed with custom MATLAB^® code (Mathworks, Natick, MA, USA).
Motion capture parameters selected for analysis sought to capture post-stroke gait, asymmetry and general apathetic behaviour observed, such as lowering of the head and shoulders and inability to extend the fetlock joint contralateral to the stroke affected hemisphere. Parameters of interest were subsequently classified into global and limb-specific. The final parameters selected for analysis and their purpose are provided in Supplementary Table S1 (global parameters) and Supplementary Table S2 (limb-specific parameters). Global parameters correspond to the outcome measures pertaining the entire trial, for example forward velocity. These outcomes were calculated as the mean value across the entire trial. Limb-specific parameters were computed from the observation of the kinematic data of each limb within its corresponding gait cycle. For each trial, the gait cycles were identified following the method from Ghoussayni et al. (43 (link)). Changes in the velocity of each limb's hoof marker (DPHAL, Figure 2) were detected in the vertical and progression directions, determining when the marker stopped moving (entering stance phase) or started moving (entering swing phase). One complete gait cycle per limb was extracted from each trial to calculate kinematic measures of interest using two-dimensional (2D) planar analysis. Planar analysis was independently performed in the vertical (sagittal) and lateral (left/right) directions. Joint angles were defined between two vectors in the sagittal plane for the fetlock, carpus, and elbow of the forelimb, and fetlock, tarsus, and stifle of the hindlimb (Figure 2).

In total, 100 weaned BDS, DBS, and DLY piglets (35 days old) were selected and raised in the same building (20 pigs per pen), equipped with a fully slatted floor, feeders, and nipple drinkers. All pigs were raised in professional breeding cooperatives in Pude Village, Malutang Township, Luquan County, Kunming City, Yunnan Province, China. The pigs were fed the same National Research Council (2012) three-stage diet (Supplementary Table 1) until 210 days of age. Ten healthy and weight-matched pigs of each breed were randomly selected. Weighed and recorded (live weight) after fasting for 24 h, then euthanized via exsanguination. Subsequently, carcass measurements were collected according to Song et al. (11 (link)) method (11 (link)), including carcass weight (remove head, hoof, tail and viscera), carcass length (the distance from the pubic symphysis leading edge to the fovea of the first cervical spine on the left side of the carcass) and lean meat rate (the percentage of lean meat weight to carcass weight). The longissimus dorsi muscle of each pig was then removed to evaluate meat quality and glycolysis potential. The samples were placed in a refrigerator at 4°C, and meat quality characteristics were evaluated after 45 min. Moreover, a section of each muscle sample was divided, packaged, and frozen at −20°C to measure lactate, glycogen, and pH at 24 and 48 h. All animal experiments were approved by the Institutional Animal Care and Use Committee of Yunnan Agricultural University (No. YNAU20211004).