Colonial-driven extinction of the blue antelope despite genomic adaptation to low population size.

Low genomic diversity is generally indicative of small population size and is considered detrimental by decreasing long-term adaptability.1,2,3,4,5,6 Moreover, small population size may promote gene flow with congeners and outbreeding depression.7,8,9,10,11,12,13 Here, we examine the connection between habitat availability, effective population size (Ne), and extinction by generating a 40× nuclear genome from the extinct blue antelope (Hippotragus leucophaeus). Historically endemic to the relatively small Cape Floristic Region in southernmost Africa,14,15 populations were thought to have expanded and contracted across glacial-interglacial cycles, tracking suitable habitat.16,17,18 However, we found long-term low Ne, unaffected by glacial cycles, suggesting persistence with low genomic diversity for many millennia prior to extinction in ∼AD 1800. A lack of inbreeding, alongside high levels of genetic purging, suggests adaptation to this long-term low Ne and that human impacts during the colonial era (e.g., hunting and landscape transformation), rather than longer-term ecological processes, were central to its extinction. Phylogenomic analyses uncovered gene flow between roan (H. equinus) and blue antelope, as well as between roan and sable antelope (H. niger), approximately at the time of divergence of blue and sable antelope (∼1.9 Ma). Finally, we identified the LYST and ASIP genes as candidates for the eponymous bluish pelt color of the blue antelope. Our results revise numerous aspects of our understanding of the interplay between genomic diversity and evolutionary history and provide the resources for uncovering the genetic basis of this extinct species' unique traits.

Agonistic interactions and social behaviors in the Saharan Dorcas gazelle (Gazella dorcas neglecta): Using social network analysis to evaluate relationships and social structure in captive male groups.

Social interactions, including agonistic behavior, are very important for the management and welfare of individuals forming groups in captivity. One of the main concerns for the stability and durability of adult male groups is a noticeable level of intraspecific aggression. This study comprises a Social Network Analysis to illustrate social structure in different groups of captive Saharan Dorcas gazelles (Gazella dorcas neglecta). Our main objectives were to examine the relationship between agonistic, affiliative, and association networks and their reciprocity, assessing also whether the agonistic networks can conform to a linear dominance hierarchy. For these purposes, we recorded the behavior of 23 adult males organized in five herds, three composed only of Saharan Dorcas gazelle males and two mixed herds in which there were also Mhorr gazelle males (Nanger dama mohor). Observations were carried out during 295 h through scan sampling. We found no correlation between the affiliative and association networks in any group, although there was a significant correlation between the agonistic and association networks in mixed-species groups which was not present in single-species groups. Overall, there was no consistent reciprocity in either affiliative nor agonistic networks and none of the agonistic networks showed a linear structure. These results indicate that affiliative behavior in Saharan Dorcas gazelles offers distinctive and valuable information about the bonds between individuals, however, their dominance structure is far more complex than previously thought. As information provided by affiliative and proximity behaviors is different in this species, we suggest considering affiliative interactions to stablish affinity between individuals. Evaluating different social behaviors and not only agonistic interactions in later studies, is also recommended to develop a more accurately daily management in zoos that guarantee group stability and individuals' welfare, which will improve the conservation of captive populations.

Diet explains Serengeti grazers' predictable migration.

Each species' dining habits set the table for the next wave.

Interplay of competition and facilitation in grazing succession by migrant Serengeti herbivores.

Competition, facilitation, and predation offer alternative explanations for successional patterns of migratory herbivores. However, these interactions are difficult to measure, leaving uncertainty about the mechanisms underlying body-size-dependent grazing-and even whether succession occurs at all. We used data from an 8-year camera-trap survey, GPS-collared herbivores, and fecal DNA metabarcoding to analyze the timing, arrival order, and interactions among migratory grazers in Serengeti National Park. Temporal grazing succession is characterized by a "push-pull" dynamic: Competitive grazing nudges zebra ahead of co-migrating wildebeest, whereas grass consumption by these large-bodied migrants attracts trailing, small-bodied gazelle that benefit from facilitation. "Natural experiments" involving intense wildfires and rainfall respectively disrupted and strengthened these effects. Our results highlight a balance between facilitative and competitive forces in co-regulating large-scale ungulate migrations.

Effects of temperament during handling and social rank on the blood biochemical parameters of common eland (Taurotragus oryx).

Large herbivores are subject to handling and social stress in captivity. These may affect blood biochemical values, which motivated this research. Twelve healthy common eland (Taurotragus oryx) were monitored for 12 months. The animals were handled monthly, and blood samples were collected. Samples from every second month were analysed for 14 blood biochemical parameters. Temperament throughout the handling, as the summation of various behavioural responses, was calculated as a proxy of the stress generated during handling. Social behaviour was recorded each month, and the agonistic interactions were used to calculate the social rank, which was considered a proxy of social stress. Generalised Linear Mixed Models were designed to test the effects of temperament and social rank on the blood biochemical parameters while keeping sex, age, body condition, and body weight as covariates. The results show that the temperament during handling influences blood levels of albumin, alkaline phosphate, blood urea nitrogen, glucose, total bilirubin, and total protein; however, social rank has little influence, affecting just albumin. The ranges observed in the values of these biochemical parameters were still within their reference intervals, implying the absence of pathology or physiological problems during the study. The results suggest that blood biochemical values of physically restrained common eland should be carefully interpreted, even in animals already habituated to routine handling. On the contrary, social rank has low effects on the blood biochemical parameters.

Movement drives population dynamics of one of the most mobile ungulates on Earth: Insights from a mechanistic model.

Long-distance movements are hypothesized to positively influence population size and stability of mobile species. We tested this hypothesis with a novel modeling approach in which moving herbivores interact with the environment created by a dynamic global vegetation model using highly mobile Mongolian gazelles in the eastern Mongolian grasslands as a case study. Gazelle population dynamics were modeled from 1901 to 2018 under two scenarios, one allowing free movement and one restricting movement. Gazelles were 2.2 times more abundant when they could move freely and were extirpated in 71% of the study area when mobility was restricted. Mobility resulted in greater population increases during times of abundant forage and smaller population decreases during drought. Reduced thermoregulatory costs associated with climate change, combined with an increase in vegetation biomass, increased gazelle abundance. Since high abundances often resulted in overgrazing and, thus, extirpation when movement was restricted, mobility had an important role in maintaining higher densities. The novel modeling approach shows how accounting for not just herbivore but also plant ecophysiology can improve our understanding of the population dynamics of highly mobile herbivores, in particular when examining the effects of habitat and climate change. Since the model simulates herbivores based on general physiological mechanisms that apply across large herbivores and the vegetation model can be applied globally, it is possible to adapt the model to other large-herbivore systems.

Rabies in kudu: Revisited.

Rabies infects all mammals; however, transmission cycles are only maintained in certain bat and carnivore species. The high incidence of rabies in Greater Kudu (Tragelaphus strepsiceros) observed in Namibia for over 40 years has led to postulation that independent virus transmission is occurring within this antelope population. We have analysed extensive experimental, epidemiological, phylogeographic and deep sequence data, which collectively refute maintenance of an independent rabies cycle in kudu. As rabies in kudu continues to have a negative impact on the Namibian agricultural sector, measures to protect kudu have been investigated, including the use of a third-generation oral rabies vaccine. Initial results show protection of kudu from rabies infection via the oral route, with an appropriate bait design, different application schedules and vaccination doses further enhancing the immune response. Rabies in kudu is a complex interplay at the wildlife-livestock interface and requires a concerted approach to successfully control.

Successful assisted rearing and reintegration of a blue wildebeest calf into a managed herd prior to weaning.

The blue wildebeest (Connochaetes taurinus; Burchell, 1823) is an African ungulate that is notoriously difficult to hand-rear and reintegrate into managed herds. The following case report is of a female wildebeest neonate that was orphaned, bottle-reared, and successfully reintroduced into a herd at a zoological facility. Since blue wildebeests are a highly precocial and herd-oriented species that calve synchronously in the wild as a reproductive strategy to maximize survival, early release of assist-reared neonates into managed herds before weaning may be an effective strategy for increasing the chance of acceptance. Wildebeest calves can also be easily trained for bottle-feedings in the field to facilitate early release. Due to a lack of literature on the assisted rearing and reintegration of wildebeest calves into managed herds, this case report provides useful techniques for the feeding, training, and releasing of bottle-reared wildebeests that may also be applicable to other assist-reared herd species at zoological facilities.

Feeding Preference and Habitat Association of Defassa Waterbuck (Kobus ellipsiprymnus defassa) in Nechisar National Park, Southern Ethiopia.

This study investigated feeding preference and habitat association of waterbucks in Nechisar National Park from November 2016 to August 2017 by total count and direct observation methods. From this survey, 20 plant species were observed to be the food items consumed by waterbucks. Waterbucks were mostly grazers than browser. The plant species consumed by waterbucks was highly significant between seasons (χ 2 = 121.34, df = 1, p < 0.05). Out of 20 total plant species consumed, annual grass (11.5%), Leersia virginica (8.4%), and Cynodon dactylon (8.4%) were the most frequently consumed food items, but Tamarindus indica (2.9%), Balanites aegyptica (3.3%), and Acacia polycantha (2.36%) were the least consumed food items. There was significant difference between plant species consumed during both seasons (χ 2 = 177.67, df = 19, p < 0.05). The Shannon-Wiener diversity index result revealed that there were different varieties of food items for waterbucks in dry season (H' = 2.9) than in wet season (H' = 2.6). Young leaves comprised the largest proportion of plant parts consumed by waterbucks compared to others. There was a highly significant difference in feeding plant parts in both seasons (χ 2 = 88.5, df = 7, p < 0.05). Waterbucks spent more time on feeding in the morning and late afternoon during both seasons. The total number of waterbucks in each habitat varied due to food availability in different seasons. Different conservation measures should be taken against waterbuck reduction and create appropriate environment for waterbuck.

Reactive anti-predator behavioral strategy shaped by predator characteristics.

Large mammalian herbivores use a diverse array of strategies to survive predator encounters including flight, grouping, vigilance, warning signals, and fitness indicators. While anti-predator strategies appear to be driven by specific predator traits, no prior studies have rigorously evaluated whether predator hunting characteristics predict reactive anti-predator responses. We experimentally investigated behavioral decisions made by free-ranging impala, wildebeest, and zebra during encounters with model predators with different functional traits. We hypothesized that the choice of response would be driven by a predator's hunting style (i.e., ambush vs. coursing) while the intensity at which the behavior was performed would correlate with predator traits that contribute to the prey's relative risk (i.e., each predator's prey preference, prey-specific capture success, and local predator density). We found that the choice and intensity of anti-predator behaviors were both shaped by hunting style and relative risk factors. All prey species directed longer periods of vigilance towards predators with higher capture success. The decision to flee was the only behavior choice driven by predator characteristics (capture success and hunting style) while intensity of vigilance, frequency of alarm-calling, and flight latency were modulated based on predator hunting strategy and relative risk level. Impala regulated only the intensity of their behaviors, while zebra and wildebeest changed both type and intensity of response based on predator traits. Zebra and impala reacted to multiple components of predation threat, while wildebeest responded solely to capture success. Overall, our findings suggest that certain behaviors potentially facilitate survival under specific contexts and that prey responses may reflect the perceived level of predation risk, suggesting that adaptive functions to reactive anti-predator behaviors may reflect potential trade-offs to their use. The strong influence of prey species identity and social and environmental context suggest that these factors may interact with predator traits to determine the optimal response to immediate predation threat.

PREGNANCY DIAGNOSIS IN OKAPI (OKAPIA JOHNSTONI) USING BIOPRYN ENZYME-LINKED IMMUNOSORBENT ASSAY FOR DETECTION OF PREGNANCY-SPECIFIC PROTEIN-B (PSPB).

Banked serum samples from seven okapi (Okapia johnstoni) with known pregnancy status were evaluated using the BioPRYN wild enzyme-linked immunosorbent assay to detect pregnancy-specific protein B (PSPB). Thirty-six serum samples, 18 from known pregnant and 18 from nonpregnant okapi, were analyzed. Using optical density cutoffs, the BioPRYN wild assay demonstrated a sensitivity of 88% (95% confidence interval, 65%-98%) and a specificity of 100% (95% confidence interval, 81%-100%). In one sample, this test confirmed pregnancy as early as 21 days of gestation; however, two pregnant okapi were reported to be not pregnant at 23 and 38 days of gestation, suggesting sensitivity may be lower in early gestation. Sensitivity improved to 100% when samples were evaluated in okapi at 116 days or greater of gestation. Analysis of PSPB can be used to augment pregnancy diagnosis in okapi, a species that is of high conservation value and has documented pregnancy-associated morbidity.

Physical characteristics of forestomach contents from two nondomestic small ruminants, the blackbuck (Antilope cervicapra) and the Arabian sand gazelle (Gazella subgutturosa marica).

Rumen content stratification and the degree of dissociation of particle and fluid retention in the reticulorumen differ between 'moose-type' and 'cattle-type' ruminant species. These differences are not strictly linked to diet, except for a seeming limitation of 'moose-type' ruminants to a browsing niche. Nevertheless, these differences can be plausibly linked to other observed differences in ruminants, such as the intraruminal papillation pattern, or the size of the omasum. However, many of the corresponding measures are still only available for a restricted number of species. Here, we investigated the dry matter (i.e., the inverse of the moisture) concentration in forestomach contents of 10 blackbuck (Antilope cervicapra) and 7 Arabian sand gazelle (Gazella subgutturosa marica), and quantified the rumen papillation pattern. The blackbucks had distinct rumen contents stratification, with more moisture in ventral than in dorsal contents (difference 3.6% units, P < 0.001), whereas this difference was much less pronounced in the sand gazelles (0.6% units, P = 0.227). While reticulum contents were particularly moist in both species, omasum contents were particularly dry in sand gazelles, but did not differ in moisture from rumen contents in the blackbuck. This species is an outlier among ruminants due to its extremely small omasum. The intraruminal papillation pattern did not differ between blackbucks and sand gazelles and showed a surface enlargement factor (SEF) in the dorsal rumen of 27-28% of the SEF in the Atrium ruminis. Compared to data on digesta retention in the same species, the findings are in line with the overall concept of a high fluid throughput causing a distinct stratification of rumen contents and intraruminal papillation, and necessitating a large omasum for fluid re-absorption. However, the data also show that individual species may not correspond to all the assumptions of the concept, suggesting taxon-specific differences between species. Reasons for these differences cannot be linked to a dietary grass-browse spectrum, but may lie in evolutionary contingency.

Long-term trends in a forest ungulate community: park establishment increases numbers, but poaching is a constant threat.

Deforestation represents one of the greatest threats to tropical forest mammals, and the situation is greatly exacerbated by bushmeat hunting. To construct informed conservation plans, information must be gathered about responses to habitat degradation, regeneration, and hunting over a sufficiently long period to allow demographic responses. We quantified changes in the abundance of three commonly occurring ungulate species (i.e., bushbuck, Tragelaphus scriptus; red duiker, Cephalophus sp.; blue duiker, Cephalophus monticola) at eight sites in Kibale National Park, Uganda (old growth=3; logged=3; regenerating=2) for 23 years. Changes in abundance (363 surveys totaling 1 450 km) were considered in regard to the park's management strategy, regional economic indicators, and estimates of illegal hunting. Bushbuck abundance increased in old-growth and logged forests from 1996 to 2009, and then oscillated around this level or declined. Duiker abundance demonstrated a similar pattern, but abundance in the old-growth forests showed a general increase from 1996 to present day. Duiker abundance in the logged forests exhibited an early increase, but subsequent oscillation. Poaching signs per patrol have remained stable over the last decade, despite increases in the size of the surrounding population, cost of living, and cost of schooling, thus reflecting successful efforts in conservation education and enforcement. Our study highlights the positive impact of park establishment, patrol, and conservation efforts on ungulate populations and shows the adaptability of forest mammal populations to different management schemes.

Optimizing livestock carrying capacity for wild ungulate-livestock coexistence in a Qinghai-Tibet Plateau grassland.

Wild ungulates are an important part of terrestrial ecosystems and play a critical role in maintaining ecosystem health and integrity. In many grassland ecosystems that are habituated by wild ungulates, the coexistence of domestic ungulates has created a conflict over grazing resources. Solving this conflict requires a balanced and sustainable policy that satisfies both the needs of wildlife protection and food production. Here, we assess the optimal grassland livestock carrying capacity of an alpine grassland on the Qinghai-Tibet Plateau, given the coexistence of wild populations of kiangs (Equus kiang) and Tibetan gazelles (Procapra picticaudata), two key species grazing in this region. We use kriging and the MaxEnt method to estimate the population sizes of kiangs and Tibetan gazelles in Maduo County, Qinghai Province. We then convert the estimated population size of the two species into sheep units and calculate the residual carrying capacity for livestock grazing. We show that after accounting for the grazing need for kiangs and Tibetan gazelles, grassland in Maduo is capable of supporting 420,641 sheep units, which is slightly more than the current livestock population. However, the residual carrying capacity is highly uneven across the region, and overgrazing is found in many areas of Maduo, especially in northern Maduo. This research provides a useful framework for planning sustainable livestock farming for the Qinghai-Tibet Plateau and other regions facing wildlife-livestock conflict.

Resource selection of a nomadic ungulate in a dynamic landscape.

Nomadic movements are often a consequence of unpredictable resource dynamics. However, how nomadic ungulates select dynamic resources is still understudied. Here we examined resource selection of nomadic Mongolian gazelles (Procapra gutturosa) in the Eastern Steppe of Mongolia. We used daily GPS locations of 33 gazelles tracked up to 3.5 years. We examined selection for forage during the growing season using the Normalized Difference Vegetation Index (NDVI). In winter we examined selection for snow cover which mediates access to forage and drinking water. We studied selection at the population level using resource selection functions (RSFs) as well as on the individual level using step-selection functions (SSFs) at varying spatio-temporal scales from 1 to 10 days. Results from the population and the individual level analyses differed. At the population level we found selection for higher than average NDVI during the growing season. This may indicate selection for areas with more forage cover within the arid steppe landscape. In winter, gazelles selected for intermediate snow cover, which may indicate preference for areas which offer some snow for hydration but not so much as to hinder movement. At the individual level, in both seasons and across scales, we were not able to detect selection in the majority of individuals, but selection was similar to that seen in the RSFs for those individuals showing selection. Difficulty in finding selection with SSFs may indicate that Mongolian gazelles are using a random search strategy to find forage in a landscape with large, homogeneous areas of vegetation. The combination of random searches and landscape characteristics could therefore obscure results at the fine scale of SSFs. The significant results on the broader scale used for the population level RSF highlight that, although individuals show uncoordinated movement trajectories, they ultimately select for similar vegetation and snow cover.

Identifying the true number of specimens of the extinct blue antelope (Hippotragus leucophaeus).

Native to southern Africa, the blue antelope (Hippotragus leucophaeus) is the only large African mammal species known to have become extinct in historical times. However, it was poorly documented prior to its extinction ~ 1800 AD, and many of the small number of museum specimens attributed to it are taxonomically contentious. This places limitations on our understanding of its morphology, ecology, and the mechanisms responsible for its demise. We retrieved genetic information from ten of the sixteen putative blue antelope museum specimens using both shotgun sequencing and mitochondrial genome target capture in an attempt to resolve the uncertainty surrounding the identification of these specimens. We found that only four of the ten investigated specimens, and not a single skull, represent the blue antelope. This indicates that the true number of historical museum specimens of the blue antelope is even smaller than previously thought, and therefore hardly any reference material is available for morphometric, comparative and genetic studies. Our study highlights how genetics can be used to identify rare species in natural history collections where other methods may fail or when records are scarce. Additionally, we present an improved mitochondrial reference genome for the blue antelope as well as one complete and two partial mitochondrial genomes. A first analysis of these mitochondrial genomes indicates low levels of maternal genetic diversity in the 'museum population', possibly confirming previous results that blue antelope population size was already low at the time of the European colonization of South Africa.

Osteomorphological features of the hind limb bones of Saiga antelope (Saiga tatarica).

The intralimb indices and calcaneal linear metrics are known as the reliable predictors of locomotor adaptation in artiodactyls. The osteological features of hindlimb in adult Saiga (Saiga tatarica) were described, and its correlation with cursoriality and habitat adaptation was discussed. Gross anatomy data showed Saiga owned the deep acetabulum as a broad lunate surface, the large acetabular anteversion, the well-developed ischiatic tuberosity and the prominent gluteal lines. It also presented the robust rough line and the strong gluteal tuberosity. A proximodistally elongated eminence located on the cranially distal tibia, which had not been found in goat. The tibial extensor groove was deep. The calcaneal tuberosity was robust. Digital anatomy data showed Saiga owned the higher metatarsal-femur ratio than forest musk deer and sheep. Comparing with wild bovids and sheep, Saiga presented a transitional variation in calcaneal form. The mean greatest length of the calcaneus (GLC) and the height of the sustentacular facet (HSF) in Saiga were shorter than that in sheep and longer than that in wild bovids respectively (F = 587.492; F = 10.264, p < .05). The wild bovids had longer cubonavicular facets than the other two groups (F = 18.587, p < .05). The great metatarsal-femur ratio of Saiga implied a superior cursorial ability and high conservation confronting the different habitats. The calcaneal linear metrics might shed light on lifestyle-related functional adaptation over decades of short-term evolution in the semi-free range environment.

Reproductive events and respective faecal androgen metabolite concentrations in captive male roan antelope (Hippotragus equinus).

Understanding the reproductive biology of the roan antelope (Hippotragus equinus) (É. Geoffroy Saint-Hilaire, 1803) is crucial to optimise breeding success in captive breeding programmes of this threatened species. In this study, the pattern of faecal androgen metabolite (fAM) production related to reproductive events (calving or birthing, mating, gestation, and lactation), sexual behaviours as well as environmental cues were studied in captive adult male roan antelope. Faecal sample collection and behavioural observations were carried out from August 2017 to July 2018 for three reproductive males participating in a conservation breeding programme at the Lapalala Wilderness Nature Reserve in South Africa. As a prerequisite, the enzyme immunoassay used in this study was biologically validated for the species by demonstrating a significant difference between fAM concentrations in non-breeding adults, breeding adults and juvenile males. Results revealed that in adults males, the overall mean fAM levels were 73% higher during the breeding period compared to the non-breeding periods, and 85% higher when exclusively compared to the lactation/gestation periods, but only 5.3% higher when compared to the birthing period. Simultaneously, fAM concentrations were lower during the wet season compared to the dry season, increasing with a reduction in photoperiod. With the exception of courtship, frequencies of sexual behaviours monitored changed in accordance with individual mean fAM concentrations in male roan antelope, the findings suggest that androgen production varies with the occurrence of mating activity and may be influenced by photoperiod but not with rainfall.

Distance sampling of duikers in the rainforest: Dealing with transect avoidance.

Bushmeat is a major source of protein and income in tropical regions but is often over-harvested. A better monitoring of bushmeat stocks could help achieve sustainability. We used a combination of simulations and transect survey data collected from blue duikers (Philantomba monticola) in the Lomako wildlife reserve, Democratic Republic of the Congo, to investigate the use of transect-based distance sampling to monitor bushmeat stocks. The comparison of dung piles and direct observations of duikers evidenced that animals avoided both the transects in the absence of observers, and the observers themselves. This type of behavioural response appeared common in a literature survey. It causes a negative bias in the estimates of population densities from the standard distance sampling methodology. This negative bias would lead to over-pessimistic predictions of population viability, especially if the behavioural response is more intense in the locations where the animals are hunted. In turn, this would lead to excessively conservative management recommendations. To correct for the effect of the behavioural response of the animals to either the transects or the observers, we recommend recording both the forward and perpendicular distances to the observers (2D distance sampling), not just the perpendicular distance. We also recommend multiple-observer protocols. As a cautionary note, we also demonstrate a scenario where the intensity of the behavioural response is too high to reliably estimate the abundance of the population. As a perspective, we outline the general principles of a local stakeholder-based program combining distance sampling with less intensive types of ecological indicators to monitor wildlife populations.

Preference of intake of different tree leaves preserved with drying and ensiling by nyala antelope (Tragelaphus angasii).

It is recommended that browse should be an important part of diets for browsing and intermediate feeding type ruminants. However, provision of browse in winter time is problematic for many zoos located in the temperate climate. Drying or freezing of branches are commonly practiced solutions to this problem, but ensiling of tree leaves could also be an attractive option. The aim of this study was to determine the effect of drying and ensiling of different tree leaves on preference of their intake by nyala antelope (Tragelaphus angasii), an intermediate feeding type ruminant. A study was conducted on five nyala fed a standard diet (meadow hay, dehydrated chopped lucerne and limited concentrates) before and during the study. Preference of intake of five feeds was tested: dried maple (A. platanoides), oak (Q. robur), willow (S. alba), and ensiled maple and lime (T. cordata) leaves. Each day two tested feeds were offered to each animal. After 4 h of presentation, feed refusals were weighed and preference ratio for each tested feed and animal was calculated. Then, preferences were ranked using a pairwise comparison chart. Dried maple and oak leaves were preferred more than other leaves whereas ensiled lime leaves were the least preferred. On the other hand, ensiled and dried maple leaves were equally preferred. Results of this study showed that palatability of ensiled tree leaves should not be a concern when feeding nyala, and likely also other browsing or intermediate feeding type ruminants, but palatability of silage may depend on the species of tree.