The South China tiger (Panthera tigris amoyensis) is a tiger subspecies unique to China and one of the top ten endangered species in the world. It used to play an important role in the overall function of the ecosystem. This study rationally screened out key prey species of the South China tiger-the Chinese serow, Chinese goral, tufted deer, water deer, Chinese muntjac, red muntjac, sambar deer, and wild boar. Candidate sites for the rewilding and reintroduction of the South China tiger were derived by exploring changes in suitable habitats for the prey using the MaxEnt model. The results show that: (1) by 2070, except for the high-suitability areas of water deer and Chinese muntjac, the areas of suitable habitats for the other six prey species would all have decreased significantly; (2) the location of the high-suitability area of the South China tiger obtained by superimposing the suitable areas of the eight prey species would be almost stable in 2050 and 2070, but the habitat index of some high- and medium-suitability areas would decrease and turn into low-suitability areas; (3) the core candidate sites were 83,415 km2 in total, of which 25,630 km2 overlapped with existing protected areas, accounting for 30.7% of the core candidate sites, and the remaining 69.3% of the core candidate sites were mostly distributed around the protected areas; (4) the maximum core candidate site area was projected to be 10,000 km2 by 2070, which could support a small population of 23 male tigers and 66 female tigers to survive and reproduce in the wild. This study revealed the core candidate sites for the rewilding of South China tigers and estimated the number of tigers that could be reintroduced to these areas, providing a preliminary research basis for promoting the rewilding of South China tigers in China.

The gut microbiota plays an important role in host nutrient absorption, immune function, and behavioral patterns. Much research on the gut microbiota of wildlife has focused on feces samples, so the microbial composition along the gastrointestinal tract of wildlife is not well reported. To address this gap, we performed high-throughput sequencing of 16s rRNA genes and ITs rRNA genes in the gastrointestinal contents of a wild adult male Chinese muntjac (Muntiacus reevesi) to comparatively analyze the microbial diversity of different gastrointestinal regions. The results showed that the dominant bacterial phyla were Firmicutes (66.19%) and Bacteroidetes (22.7%), while the dominant fungal phyla were Ascomycetes (72.81%). The highest bacterial diversity was found in the stomach, and the highest fungal diversity was found in the cecum. The microbial communities of the large intestine and small intestine were of similar structures, which were distinct from that of the stomach. These results would facilitate the continued exploration of the microbial composition and functional diversity of the gastrointestinal tract of wild Chinese muntjacs and provide a scientific basis for microbial resource conservation of more wildlife.

Variation in the chromosome numbers can arise from the erroneous mitosis or fusion and fission of chromosomes. While the mitotic errors lead to an increase or decrease in the overall chromosomal substance in the daughter cells, fission and fusion keep this conserved. Variations in chromosome numbers are assumed to be a crucial driver of speciation. For example, the members of the muntjac species are known to have very different karyotypes with the chromosome numbers varying from 2n=70+3B in the brown brocket deer to 2n=46 in the Chinese muntjac and 2n=6/7 in the Indian muntjac. The chromosomal content in the nucleus of these closely related mammals is roughly the same and various chromosome fusion and fission pathways have been suggested as the evolution process of these karyotypes. Similar trends can also be found in lepidoptera and yeast species which show a wide variation of chromosome numbers. The effect of chromosome number variation on the spindle assembly time and accuracy is still not properly addressed. We computationally investigate the effect of conservation of the total chromosomal substance on the spindle assembly during prometaphase. Our results suggest that chromosomal fusion pathways aid the microtubule-driven search and capture of the kinetochore in cells with monocentric chromosomes. We further report a comparative analysis of the site and percentage of amphitelic captures, dependence on cell shape, and position of the kinetochore in respect to chromosomal volume partitioning.

The family Cervidae is the second most diverse family in the infraorder Pecora and is characterized by a striking variability in the diploid chromosome numbers among species, ranging from 6 to 70. Chromosomal rearrangements in Cervidae have been studied in detail by chromosome painting. There are many comparative cytogenetic data for both subfamilies (Cervinae and Capreolinae) based on homologies with chromosomes of cattle and Chinese muntjac. Previously it was found that interchromosomal rearrangements are the major type of rearrangements occurring in the Cervidae family. Here, we build a detailed chromosome map of a female reindeer (Rangifer tarandus, 2n = 70, Capreolinae) and a female black muntjac (Muntiacus crinifrons, 2n = 8, Cervinae) with dromedary homologies to find out what other types of rearrangements may have underlined the variability of Cervidae karyotypes. To track chromosomal rearrangements and the distribution of nucleolus organizer regions not only during Cervidae but also Pecora evolution, we summarized new data and compared them with chromosomal maps of other already studied species. We discuss changes in the pecoran ancestral karyotype in the light of new painting data. We show that intrachromosomal rearrangements in autosomes of Cervidae are more frequent than previously thought: at least 13 inversions in evolutionary breakpoint regions were detected.

Abstract AimGlobal warming is threatening species survival worldwide. Cold‐adapted montane ungulates may be among the first species to be affected by global warming. Here, we investigate the behavioural thermoregulation hypothesis that montane ungulates buffer the effects of warming climate through behavioural adjustment.LocationFoping National Nature Reserve (FNNR) on the southern slopes of the Qinling Mountains, Shaanxi, China.MethodsWe used infrared camera traps over six consecutive years to monitor behavioural responses of montane ungulates to seasonal and diurnal changes in ambient temperature. We evaluated how seasonal altitudinal migration and changes to their daily activity budget may mitigate the effect of warming on montane ungulates.ResultsAll four montane ungulate species examined here preferred ambient temperatures <22°C in summer. On this basis, we estimate that 27.9% (+2°C) and 46.7% (+3°C) of suitable summer habitats will become unsuitable under climate change. Chinese muntjac and tufted deer displayed behavioural changes to daily activity budgets, which may help them overcome global climate warming to a certain extent. By contrast, golden takin and wild boar did not display similar behavioural changes, and it may be because they are not affected by current shifts in climate due to larger body size and thus lower thermal stress.Main ConclusionsThese analyses demonstrate different species responses to warming are likely and are determined by specific tolerances to ambient temperature thresholds. However, it remains to be seen whether species interactions caused by behavioural shifts may affect species ability to respond to climate.

IntroductionOver recent decades, the abundance and geographic ranges of wild ungulate species have expanded in many parts of Europe, including the UK. Populations are managed to mitigate their ecological impacts using interventions, such as shooting, fencing and administering contraception. Predicting how target species will respond to interventions is critical for developing sustainable, effective and efficient management strategies. However, the quantity and quality of evidence of the effects of interventions on ungulate species is unclear. To address this, we systematically mapped research on the effects of population management on wild ungulate species resident in the UK.MethodsWe searched four bibliographic databases, Google Scholar and nine organisational websites using search terms tested with a library of 30 relevant articles. Worldwide published peer-reviewed articles were considered, supplemented by ‘grey’ literature from UK-based sources. Three reviewers identified and screened articles for eligibility at title, abstract and full-text levels, based on predefined criteria. Data and metadata were extracted and summarised in a narrative synthesis supported by structured graphical matrices.ResultsA total of 123 articles were included in the systematic map. Lethal interventions were better represented (85%, n = 105) than non-lethal interventions (25%, n = 25). Outcomes related to demography and behaviour were reported in 95% of articles (n = 117), whereas effects on health, physiology and morphology were studied in only 11% of articles (n = 14). Well-studied species included wild pigs (n = 58), red deer (n = 28) and roe deer (n = 23).ConclusionsEvidence for the effects of population management on wild ungulate species is growing but currently limited and unevenly distributed across intervention types, outcomes and species. Priorities for primary research include: species responses to non-lethal interventions, the side-effects of shooting and studies on sika deer and Chinese muntjac. Shooting is the only intervention for which sufficient evidence exists for systematic review or meta-analysis.

Biodiversity monitoring is an important means by which to evaluate management effectiveness and develop sound conservation plans. In this study, 52 cameras were installed in the study area of Tianzhushan to assess wildlife diversity and elevational patterns from 2018 to 2019. In total, 9 541 independent photos were collected within 26 565 camera-days. We analysed the relative abundance index (RAI), activity patterns and seasonal activity rhythms of the five most abundant species at 200-m elevational intervals throughout the study area (elevation range 1 400–2 000 m a.s.l.). Based on RAI results, the activity patterns of the five most abundant species (i.e. Susscrofa, Naemorhedusgriseus, Muntiacusreevesi, Arctonyxcollaris and Capricornismilneedwardsii) were analysed. Amongst the detected wild mammals, Susscrofa had the highest RAI value of 9.91, while the occurrence of Tamiopsswinhoei had the lowest RAI at 0.004. In addition, there were significant differences in species activity during the daytime and night-time. RAI of the ungulate community peaked at the 1 600–1 800 m a.s.l. elevational range, thus showing a mid-elevational peak pattern. Differences in vertical distribution patterns were detected for the five most abundant species. Mainland serows and wild boars preferred mid-elevation habitats (1 600–1 800 m a.s.l.), long-tailed gorals preferred mid- and high altitudes (1 600–2 000 m a.s.l.) elevation habitats and Chinese muntjacs and hog badgers preferred low elevation habitats (1 400–1 600 m a.s.l.). Taken together, this is the first study to reveal the five dominant species activity patterns in Tianzhushan, which is of importance for wildlife conservation.

The competitive relationship and coexistence pattern among close related species have long been one of the hot issues in ecological research. Interspecies interactions can exert important influences on the local distribution of rare species. Black muntjac Muntiacus crinifrons is an endemic species to eastern China, currently restricted to limited regions. In contrast, Chinese muntjac Muntiacus reevesi is the most common and widespread deer in southern China. Both species co‐occur in southern Anhui and western Zhejiang Province. Little is known about the interaction of these two sympatric‐related species. In this study, to investigate the site use determinants and co‐occurrence pattern of the two sympatric muntjac species, we conducted a camera trap survey across about 250 km2 in mountainous area of southern Anhui Province, China. We adopted a multistep approach to incorporate habitat preferences while modeling occupancy and detection. We found that the two species did not separate along elevation gradient (range from 400 m to 1,400 m) as described in previous studies. Results of single‐species occupancy models indicated that elevation had positive effects on the site use of both species, while slope had an opposite influence on their site use. Positive effects of elevation on the site use implied that both species try to avoid human interference at low elevations. Significant negative effect of slope on the site use of black muntjac suggested that the species prefer habitat with gentle slope and avoided steep. Co‐occurrence models and species interaction factors provided evidence that the two muntjac species had an independent occupancy (ψ BM CM = ψ BM cm, SIF = 1) and exhibited a positive species interaction in detection probability (p BM < r BM CM). Combined with the results of previous studies, we suggested that it was fine differentiation in microhabitats and food resources utilization rather spatial or temporal segregation that allowed the two species co‐occurrence. The site use determinants revealed in our study would be useful for the habitat conservation and restoration for the rare black muntjac, and the co‐occurrence pattern of the two sympatric muntjac species could provide useful information for deep understanding of the coexistence mechanism among forest‐dwelling ungulates.

Climate fluctuations in the past and in the future are likely to result in population expansions, shifts, or the contraction of the ecological niche of many species, and potentially leading to the changes in their geographical distributions. Prediction of suitable habitats has been developed as a useful tool for the assessment of habitat suitability and resource conservation to protect wildlife. Here, we model the ancestral demographic history of the extant modern Chinese Muntjac Muntiacus reevesi populations using approximate Bayesian computation (ABC) and used the maximum entropy model to simulate the past and predict the future spatial dynamics of the species under climate oscillations. Our results indicated that the suitable habitats for the M. reevesi shifted to the Southeast and contracted during the Last Glacial Maximum, whereas they covered a broader and more northern position in the Middle Holocene. The ABC analyses revealed that the modern M. reevesi populations diverged in the Middle Holocene coinciding with the significant contraction of the highly suitable habitat areas. Furthermore, our predictions suggest that the potentially suitable environment distribution for the species will expand under all future climate scenarios. These results indicated that the M. reevesi diverged in the recent time after the glacial period and simultaneously as its habitat’s expanded in the Middle Holocene. Furthermore, the past and future climate fluctuation triggered the change of Chinese muntjac spatial distribution, which has great influence on the Chinese muntjac’s population demographic history.

Ruminant methane, which is generated by methanogens through the consumption of hydrogen and supports the normal function of the rumen ecosystem, is a major source of greenhouse gases. Reductive acetogenesis by acetogens is a possible alternative sink that can dispose of hydrogen for acetate production. However, the distribution of rumen methanogens and acetogens along with the relationships among methanogens, acetogens, and their host are poorly understood. Therefore, we investigated the rumen methanogen and acetogen communities of 97 individual animals representing 14 ruminant species within three ruminant families Cervidae (deer), Bovidae (bovid), and Moschidae (musk deer). The results showed that the Methanobrevibacter spp. and acetogens associated with Eubacteriaceae were the most widespread methanogens and acetogens, respectively. However, other methanogens and acetogens exhibited host specificity in the rumen of reindeer and Chinese muntjac deer. Acetogen and methanogen communities were not correlated in these species, and the phylosymbiosis signature between host phylogeny and the composition of both communities was lacking. The abundance of Methanobrevibacter gottschalkii was negatively correlated with the degree of papillation of the rumen wall. Finally, co-occurrence analysis showed that the variation of the predicted methane yields was characterized by the interactive patterns between methanogens, acetogens, and concentrations of rumen metabolites. Our results show that rumen methanogen and acetogen communities have low compositional interdependence and do not exhibit parallel host evolution, which suggests that the strategies for mitigating methane production should be based on a species-specific rumen microbiota analysis.

Isolation and culture of somatic cells from animals especially endangered species have raised great concerns as it is being an effective and convenient way to preserve genetic materials for future studies. As a species native to China, Chinese muntjac (Muntiacus reevesi) is listed as a beneficial species with economic and scientific research values. To our knowledge, however, there have been no published reports on somatic cell preservation of this species to date. To conserve biological resources for sustainability of Chinese muntjacs' genetic diversity, we established a fibroblast cell line from the postmortem ear skin of an adult male Chinese muntjac. The cultured cells were adherent to the plastic and showed an elongated, thin, and spindle-like shape. Moreover, they were FSP1- and VIM-positive characterizing them to be fibroblastic. No microorganisms (bacteria, fungi, or mycoplasmas) were detected throughout the whole study. Cell viability was high although it declined somehow after passaging. The population doubling time was 21.28h according to the growth curve. Chromosome analysis revealed that the established fibroblast cell line contained 23 pairs of chromosomes, one pair of which was sex chromosomes (XY). Mitochondrial cytochrome C oxidase I gene of cultured cells shared 98.32% identity with those of Muntiacus reevesi registered in GenBank, which verified the cell line was derived from Muntiacus reevesi. In conclusion, we propagated and characterized fibroblast cells from a Chinese muntjac. We believe that this somatic cell line could facilitate animal cloning and breeding studies and become a useful in vitro model to address genetic questions.

Abstract AimThe characterization of the genetic structure and demographic dynamics of a species can shed light on the impacts of natural landscapes, historical climate change and human activities, thus enabling us to develop appropriate protection and management strategies. In this study, we aimed to characterize how the above‐mentioned factors shaped the genetic structure and demographic history of the Chinese muntjac (Muntiacus reevesi).LocationThe reaches of the Yangtze River, China.MethodsUsing mtDNA control region sequences and 15 nuclear microsatellite loci as genetic markers, we analysed population genetic structure with a factorial correspondence analysis (FCA), AMOVA,FSTestimates and Bayesian cluster analysis. In addition, we estimated the demographic history using Bayesian skyline plot (BSP) analysis and MSVAR simulation.ResultsHigh genetic diversity was detected in Chinese muntjac populations. Population structure analysis strongly suggesting three genetic clusters (QL/BA/WLS, DB and JLS/WN/WYS). The demographic analyses showed that Chinese muntjac experienced population fluctuation during the late Pleistocene, followed by bottleneck during the Holocene.Main conclusionsThe extant Chinese muntjac populations harbour a surprisingly high genetic diversity despite of the bottleneck they went through. The populations show significant population structure separating populations in the upper, middle and lower regions of the Yangtze River, supporting the presence of three independent management units (QL/BA/WLS, DB and JLS/WN/WYS). The population demography of the Chinese muntjac presents high concordance with the rotation of glacial–interglacial in late Pleistocene. In addition, our study identified that the Chinese muntjac's recent population history may have been influenced by early human activities (8,509–1,605 years ago).

基于粪便DNA的小麂亲权鉴定和婚配制研究

The aim of the article was to describe the pattern of main arteries at the encephalon base, their connections, and varieties. This study included 106 specimens of the head and cerebral arteries of the following eight species of the cervid family: reindeer, chital, Eld's deer, wapiti, sika deer, fallow deer, Pere David's deer or milu, and Reeve's or Chinese muntjac. The arteries of the animals under study were filled with acetone-dissolved stained vinyl superchloride or stained latex LBS3060. The analysis of the specimens revealed that the vascular system of the species of the studied cervid family was similar to the system described in other ruminant species. A branch diverging from the condylar artery to the rostral epidural rete mirabile is present in all studied cervids, but it can be found also in giraffe and eland. The pattern of the arterial vascularization of the encephalon base, which we observed in our analysis, confirms the morphological similarity between those animals and the correct placement of the cervid family in species taxonomy.

Fourteen novel polymorphic microsatellite markers were isolated and characterized for Muntiacus reevesi, a small deer species. An (AG)n enriched library was created from two individuals following the FIASCO protocol (Fast Isolation by AFLP of Sequences COtaining repeats). 14 primers were designed from 98 microsatellite sequences and tested in 32 samples. The number of alleles per locus ranged from 2 to 13 and the expected heterozygosities from 0.123 to 0.916. The loci had an average polymorphic information content value of 0.676. Five loci showed significant deviations from Hardy–Weinberg equilibrium and no linkage disequilibrium was detected. These markers should be a useful tool for further population genetic studies of Muntiacus reevesi.

Defence formation in the captive group of golden takin (Budorcas bedfordi)

Five satellite DNA families (designated as satellite I–V) have been identified in the Cervidae so far. Among those, satellite I, II and IV are centromere specific. Satellite I and II are shared by large number of deer species, where satellite IV is highly conserved among several deer species examined. Satellite III was initially thought to be roe deer specific but later identified in Chinese water deer as well. SatelliteV is Y-chromosome specific for several Asian deer species examined but also found in the pericentric region of Indian muntjac chromosome 3 and in X chromosome of Chinese water deer. The observation of interstitial hybridization sites on Indian muntjac chromosomes with satellite DNA I probe generated from Chinese muntjac provides the first molecular evidence supporting the tandem fusion theory that 2n=6♀/7♂of Indian muntjac karyotype could derive from an ancestral Chinese muntjac-like species with 2n=46. Interspecies chromosome painting study and the maximum number of interstitial hybridization detected with satellite I and satellite II DNA probes lend support to the hypothesis that the Indian muntjac karyotype could evolve directly from an ancestral Chinese water deer-like species with 2n=70. Such hypothesis is further substantiated by the finding of satellite V signals presented in specific chromosome regions between the Chinese water deer and the Indian muntjac chromosomes.

The muntjacs (Muntiacus, Cervidae) have been extensively studied in terms of chromosomal and karyotypic evolution. However, little is known about their meiotic chromosomes particularly the recombination patterns of homologous chromosomes. We used immunostained surface spreads to visualise synaptonemal complexes (SCs), recombination foci and kinetochores with antibodies against marker proteins. As in other mammals pachytene was the longest stage of meiotic prophase. 39.4% of XY bivalents lacked MLH1 foci compared to less than 0.5% of autosomes. The average number of MLH1 foci per pachytene cell in M. reevesi was 29.8. The distribution of MLH1 foci differed from other mammals. On SCs with one focus, the distribution was more even in M. reevesi than in other mammals; for SCs that have two or more MLH1 foci, usually there was a larger peak in the sub-centromere region than other regions on SC in M. reevesi. Additionally, there was a lower level of interference between foci in M. reevesi than in mouse or human. These observations may suggest that the regulation of homologous recombination in M. reevesi is slightly different from other mammals and will improve our understanding of the regulation of meiotic recombination, with respect to recombination frequency and position.

Genome-wide homologies between the tsessebe (Damaliscus lunatus, 2n = 36) and Chinese muntjac (Muntiacus reevesi, 2n = 46) have been established by cross-species painting with Chinese muntjac chromosome paints. Twenty-two autosomal painting probes detected 35 orthologous segments in the tsessebe. Hybridization results confirmed that: (i) D. lunatus carries the (9;14) reciprocal translocation that has been proposed to be a derived chromosomal landmark shared by all species of the Antilopinae; (ii) the karyotype of D. lunatus can be derived almost exclusively from the bovid ancestral karyotype through 12 Robertsonian translocations involving 24 ancestral acrocentric autosomes; (iii) in addition to the Rb fusions, pericentric heterochromatic amplification has shaped the morphology of several of the D. lunatus chromosomes. Integrated analysis of these and published cytogenetic data on pecorans has allowed us to accurately discern the karyotype history of Damaliscus (D. lunatus; D. pygargus, 2n = 38; D. hunteri, 2n = 44). The phylogenomic relationships of 3 species reflected by specific chromosomal rearrangements were consistent with published phylogenies based on morphology, suggesting that chromosomal rearrangements have played an important role in speciation within the Alcelaphini, and that karyotype characters are valuable phylogenetic markers in this group.

Considering the giraffe (Giraffa camelopardalis, GCA, 2n = 30) as a primitive species, its comparative genomic data are critical for our understanding of the karyotype evolution of pecorans. Here, we have established genome-wide chromosomal homologies between giraffe, Chinese muntjac (Muntiacus reevesi, MRE, 2n = 46) and human (Homo sapiens, HSA, 2n = 46) with whole sets of chromosome-specific paints from Chinese muntjac and human, in addition to providing a high-resolution G-banding karyotype of giraffe. Chinese muntjac and human chromosome paints detected 32 and 45 autosomal homologs in the genome of giraffe, respectively. Our results suggest that it would require at least thirteen fissions, six fusions and three intrachromosomal rearrangements to ‘transform’ the 2n = 44 eutherian ancestral karyotype to the 2n = 58 pecoran ancestral karyotype. During giraffe evolution, some ancestral eutherian syntenies (i.e. association of HSA3/21, 4/8, 7/16, 14/15, 16/19 and two forms of 12/22) have been retained, while several derived syntenies (i.e. associations of human homologous segments 2/1, 2/9, 5/19, 4/12/22, 8/9, and 10/20) have been produced. The reduction of chromosome number in giraffe from the 2n = 58 pecoran ancestral karyotype could be primarily attributed to extensive Robertsonian translocations of ancestral chromosomal segments. More complex chromosomal rearrangements (including tandem fusion, centromere repositioning and pericentric inversion) have happened during the evolution of GCA2 and GCA8.