The comparative anatomy of the petrosal bone and bony labyrinth of four small-sized deer.

Here we provide complete 3D reconstructions of the petrosal bone and bony labyrinth of four kinds of small-sized deer (Elaphodus cephalophus, Muntiacus reevesi, Muntiacus muntjak, Hydropotes inermis) based on high-resolution CT scanning, and select one musk deer (Moschus moschiferus) as a comparative object. The petrosal bone and bony labyrinth of E. cephalophus are illustrated for the first time, as well as the petrosal bones of M. reevesi and H. inermis. Some morphological characters of petrosal bone and bony labyrinth can be used to distinguish the above-mentioned species. For example, M. moschiferus shows a prominent transpromontorial sulcus and a ventral basicapsular groove on the petrosal bone; there is a bifurcate cochlear aqueduct on the bony labyrinth of E. cephalophus; there is a distinct fusion between the lateral and posterior semicircular canals on the bony labyrinth of H. inermis. Meanwhile, there are some intraspecific variations on the subarcuate fossa, the tegmen tympani, the cochlear aqueduct, as well as the endolymphatic sac. Our results further confirm that the petrosal bone and bony labyrinth have enormous potential for taxonomy. This work will provide new anatomical data for the phylogenetic study of ruminants in the future, and it will be very practical to identify the isolated ruminants' petrosal bones that are frequently unearthed from paleontological or archeological sites.

Hi-C sequencing unravels dynamic three-dimensional chromatin interactions in muntjac lineage: insights from chromosome fusions in Fea's muntjac genome.

Eukaryotes have varying numbers and structures of characteristic chromosomes across lineages or species. The evolutionary trajectory of species may have been affected by spontaneous genome rearrangements. Chromosome fusion drastically alters karyotypes. However, the mechanisms and consequences of chromosome fusions, particularly in muntjac species, are poorly understood. Recent research-based advancements in three-dimensional (3D) genomics, particularly high-throughput chromatin conformation capture (Hi-C) sequencing, have allowed for the identification of chromosome fusions and provided mechanistic insights into three muntjac species: Muntiacus muntjak, M. reevesi, and M. crinifrons. This study aimed to uncover potential genome rearrangement patterns in the threatened species Fea's muntjac (Muntiacus feae), which have not been previously examined for such characteristics. Deep Hi-C sequencing (31.42 × coverage) was performed to reveal the 3D chromatin architecture of the Fea's muntjac genome. Patterns of repeated chromosome fusions that were potentially mediated by high-abundance transposable elements were identified. Comparative Hi-C maps demonstrated linkage homology between the sex chromosomes in Fea's muntjac and autosomes in M. reevesi, indicating that fusions may have played a crucial role in the evolution of the sex chromosomes of the lineage. The species-level dynamics of topologically associated domains (TADs) suggest that TAD organization could be altered by differential chromosome interactions owing to repeated chromosome fusions. However, research on the effect of TADs on muntjac genome evolution is insufficient. This study generated Hi-C data for the Fea's muntjac, providing a genomic resource for future investigations of the evolutionary patterns of chromatin conformation at the chromosomal level.

Microtubule search-and-capture model evaluates the effect of chromosomal volume conservation on spindle assembly during mitosis.

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.

Fluid and particle retention in a small New World and a small Old World cervid, the southern pudu (Pudu puda) and Reeves's muntjac (Muntiacus reevesi).

Ruminants differ in the pattern how small particles and liquids pass through their gastrointestinal tract, and in particular their reticulorumen (RR). Based on that they may be classified into 'moose-type' and 'cattle-type' species (smaller and larger differences between particle and liquid passage, respectively). The ratio between the retention of particles and fluids is called the 'selectivity factor' (SF) and is a species-specific characteristic, studied in tragulids, giraffids and bovids, but not in many cervid species. Recently, it has been suggested that a high SF might also serve to wash digesta clean of external abrasives prior to regurgitation for rumination. In this study, we measured SF and passage kinetics (using a liquid marker and markers of different particle size, fed with the diet) in a capreoline deer, the southern pudu (Pudu puda, n = 5, 10.3 ± 2.9 kg, kept at two zoos) and a cervine deer, the Reeves's muntjac (Muntiacus reevesi, n = 6, 11.0 ± 1.7 kg, kept at a research facility). The relative daily dry matter intake (38 ± 3 g/kg0.75 for pudu and 76 ± 5 g/kg0.75 for muntjac) was higher, and the mean retention times (MRT) correspondingly shorter (e.g., MRT small particles in the total digestive tract 39 ± 8 h for pudu and 15 ± 2 h for muntjac), in the muntjac. The SF for small particles/liquid in the reticulorumen were, however, similar for both species, at 1.47 ± 0.21 for pudu and 1.66 ± 0.20 for muntjac, indicating a 'moose-type' physiology for both, irrespective of their different phylogenetic origin. To date, SF recorded in bovids attain distinctively higher values than the few reported for cervids. This situation reflects the degree of hypsodonty (tooth crown height) attained by these taxa, which is higher in bovids than in cervids. Together, constraints in hypsodonty as well as SF might limit cervids to more mesic habitats without distinct loads of external abrasives (such as dust or grit) on their food. In both species, some animals showed the typical ruminant pattern of a longer MRT for large than for small particle markers, but in some animals, this difference was not evident. This may be due to variable degrees of marker chewing during ingestion.

Comparative studies of X chromosomes in Cervidae family.

The family Cervidae is the second most diverse in the infraorder Pecora and is characterized by variability in the diploid chromosome numbers among species. X chromosomes in Cervidae evolved through complex chromosomal rearrangements of conserved segments within the chromosome, changes in centromere position, heterochromatic variation, and X-autosomal translocations. The family Cervidae consists of two subfamilies: Cervinae and Capreolinae. Here we build a detailed X chromosome map with 29 cattle bacterial artificial chromosomes of representatives of both subfamilies: reindeer (Rangifer tarandus), gray brocket deer (Mazama gouazoubira), Chinese water deer (Hydropotes inermis) (Capreolinae); black muntjac (Muntiacus crinifrons), tufted deer (Elaphodus cephalophus), sika deer (Cervus nippon) and red deer (Cervus elaphus) (Cervinae). To track chromosomal rearrangements during Cervidae evolution, we summarized new data, and compared them with available X chromosomal maps and chromosome level assemblies of other species. We demonstrate the types of rearrangements that may have underlined the variability of Cervidae X chromosomes. We detected two types of cervine X chromosome-acrocentric and submetacentric. The acrocentric type is found in three independent deer lineages (subfamily Cervinae and in two Capreolinae tribes-Odocoileini and Capreolini). We show that chromosomal rearrangements on the X-chromosome in Cervidae occur at a higher frequency than in the entire Ruminantia lineage: the rate of rearrangements is 2 per 10 million years.

Optimized protocol for live-cell analysis of kinetochore fiber maturation in Indian muntjac cells.

Here, we take advantage of the low chromosome number (2N=6) and distinctively large kinetochores of female Indian muntjac cells to investigate the molecular mechanism underlying k-fiber maturation. We describe steps for monitoring kinetochore-microtubule dynamics over time. Specifically, we detail the combination of live-cell super-resolution CH-STED microscopy of microtubule growth events within individual k-fibers and a laser-mediated k-fiber injury/repair assay. These tools provide a direct assessment of microtubule amplification mechanisms within k-fibers in metazoans. For complete details on the use and execution of this protocol, please refer to Almeida et al. (2022).1.

Genetic similarities and phylogenetic analysis of Muntjac (Muntiacus spp. ) by comparing the nucleotide sequence of 16S rRNA and cytochrome B genome / Semelhanças genéticas e análise filogenética de muntjac (Muntiacus spp. ) comparando a sequência de nucleótidos de rRNA 16S e genoma citocromo B

This study aimed to identify the phylogenetic similarities among the muntjac (Muntiacus spp.). The phylogenetic similarities among seven major muntjac species were studied by comparing the nucleotide sequence of 16s rRNA and cytochrome b genome. Nucleotide sequences, retrieved from NCBI databases were aligned by using DNASTAR software. A phylogenetic tree was created for the selected species of muntjac by using the maximum likelihood method on MEGA7 software. The results of nucleotide sequences (16s rRNA) showed phylogenetic similarities between, the M. truongsonensis and M. rooseveltorum had the highest (99.2%) while the lowest similarities (96.8%) found between M. crinifrons and M. putaoensi. While the results of nucleotide sequences (Cty b) showed the highest similarity (100%) between M. muntjak and M. truongsonensis and the lowest s (91.5%) among M. putaoensis and M. crinifrons. The phylogenetic tree of muntjac species (16s rRNA gene) shows the main two clusters, the one including M. putaoensis, M. truongsonensis, M. rooseveltorum, and M. muntjak, and the second one including M. crinifrons and M. vuquangensis. The M. reevesi exists separately in the phylogenetic tree. The phylogenetic tree of muntjac species using cytochrome b genes shows that the M. muntjak and M. truongsonensis are clustered in the same group.

Este estudo visou identificar as semelhanças filogenéticas entre os muntjac (Muntiacus spp.). As semelhanças filogenéticas entre sete grandes espécies muntjac foram estudadas comparando a sequência de nucleótidos de 16s rRNA e genoma citocromo b. As sequências de nucleótidos, obtidas a partir de bases de dados NCBI, foram alinhadas utilizando o software DNASTAR. Foi criada uma árvore filogenética para as espécies selecionadas de muntjac utilizando o método de probabilidade máxima no software MEGA7. Os resultados das sequências de nucleótidos (16s rRNA) mostraram semelhanças filogenéticas entre o M. truongsonensis e o M. rooseveltorum tiveram o maior número (99,2%) enquanto as semelhanças mais baixas (96,8%) encontradas entre M. crinifrons e M. putaoensi. Enquanto os resultados das sequências de nucleótidos (Cty-b) apresentaram a maior semelhança (100%) entre M. muntjak e M. truongsonensis e os mais baixos (91,5%) entre M. putaoensis e M. crinifrons. A árvore filogenética das espécies muntjac (gene rRNA 16s) mostra os dois principais aglomerados, o que inclui M. putaoensis, M. truongsonensis, M. rooseveltorum e M. muntjak, e o segundo incluindo M. crinifrons e M. vuquangensis. O M. reevesi existe separadamente na árvore filogenética. A árvore filogenética das espécies muntjac usando genes citocromo b mostra que os M. muntjak e M. truongsonensis estão agrupados no mesmo grupo.

Comparative Studies of Karyotypes in the Cervidae Family.

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.

Increased intake of mono- and disaccharides by Reeves's muntjac (Muntiacus reevesi). Effect on gastrointestinal tract structure and function and blood parameters.

The aim of this study was to determine the effect of an increased mono- and disaccharide (MD) intake on selected functions and structure of the gastrointestinal tract (GIT), and selected blood parameters in Reeves's muntjac (Muntiacus reevesi), a small browsing ruminant. Eighteen male muntjacs were fed diets consisting of lucerne (ad libitum), a high fibre pellet (100 g/day) and wheat bran (30 g/day) without (MD0) or with addition of 10 or 20 g of glucose, fructose and sucrose mixture/day (MD10 and MD20, respectively) for 14 days. MD dosages were set to increase intake of these saccharides by 25% and 50% relative to MD0, which resulted in a range of water-soluble carbohydrate content in the consumed dry matter from 7% to 12%. Compared to MD0 animals, MD20 animals had a lower dry matter intake, a higher MD concentrations in the reticulorumen (RR), abomasal and small intestinal digesta, higher ruminal butyrate concentration, higher SGLT1 expression in the epithelium of proximal jejunum, higher plasma glucose, lower RR tissue weight but greater caecal tissue weight (p ≤ 0.05), and had or tended to have shorter papillae and lower mucosa surface area in the Atrium ruminis (by 44%; p = 0.02 and p = 0.10, respectively); MD10 animals tended to have higher MD concentrations in the abomasal and small intestinal digesta (p ≤ 0.10), and a higher amylolytic activity (p = 0.02) as well as a tendency to lower xylanolytic activity in the RR digesta (p = 0.06). MD supplementation did not affect ruminal pH. In conclusion, low to moderate increase of MD intake increased MD concentrations in the RR, abomasal and intestinal digesta, and SGLT1 expression in intestinal epithelium, suggesting incomplete fermentation of those saccharides in the RR. MD supplementation dose-dependently affects structure of GIT in Reeves's muntjac.

Effect of sugar and starch supplementation on feed intake and nutrient digestibility in addax (Addax nasomaculatus) and Reeves's muntjac (Muntiacus reevesi).

Two studies were conducted to determine the effect of the supplementation of sugar, starch or both on feed, nutrient and energy intake and total tract digestibility in four adult female addax (Addax nasomaculatus) and four adult male Reeves's muntjac (Muntiacus reevesi) - representatives of grazing and browsing ruminants, respectively. Studies in both species were conducted according to 4 × 4 Latin Square Design. Animals had free access to meadow hay (addax) or dehydrated chopped lucerne (muntjac), and were fed a restricted amount of a 'basal diet' consisting of: (1) wheat bran; (2) wheat bran and sucrose (source of sugar); (3) wheat bran and wheat (source of starch); or (4) wheat bran, sucrose and wheat. The amounts of supplemental sucrose and wheat were set to account for 2% and 15%, respectively, of dry matter (DM) consumed. There was no effect of the ~2% sugar supplementation on DM intake of hay by addax, while the ~10% starch supplementation reduced DM intake of hay by 13% (p < 0.01); total DM intake (of hay and the basal diet) was not affected neither by sugar nor starch supplementation. When the diet for addax included wheat, this resulted in a greater intake of crude protein by 15%, lower intake of ADF by 9%, and greater crude protein digestibility by 10% (p ≤ 0.05). The ~2% sugar supplementation did not affect intake of lucerne and total DM intake by muntjac, but the ~10% starch supplementation decreased DM intake of lucerne by 25% (p < 0.01), total DM intake by 7% (p = 0.02) and intake of all nutrients (p ≤ 0.10). In summary, if high intake of roughages by captive ruminants is fundamental for their gastrointestinal functions and health, then starchy feeds supplementation should be limited, as they have an especially negative impact on roughage intake.

A comparative study on the morphology, radiography and computed tomography of the skull bones of barking deer (Muntiacus muntjak) and sambar deer (Rusa unicolor).

BACKGROUND: There is scanty information on the skull morphology of barking and sambar deer; thus the present study was designed to provide information on morphology, radiography and computed tomography (CT) of the skull bones of both deer species. MATERIALS AND METHODS: The study was conducted on 12 skulls of adult barking deer (n = 6) and sambar deer (n = 6) of either sex (n = 3 males and n = 3 females) collected from Aizawl Zoological Park, Aizawl, Mizoram. The skulls of both species were macerated as per the standard maceration techniques. RESULTS: The skull bones of both deer species were divided into a neurocranium and a viscerocranium. The neurocranium was comprised of occipital, sphenoid, temporal, frontal, parietal, interparietal and ethmoid bones. The viscerocranium consisted of nasal, lacrimal, zygomatic, maxilla, incisive, palatine, pterygoid, vomer, mandible, turbinates and hyoid bones. The cranial cavity was oval and elongated caudally. The orbit was round, complete in barking deer; however, it was oval, complete in sambar deer. The facial tuberosity was present caudal to infraorbital foramen and dorsally at superior third premolar tooth in barking deer whereas dorsally at the superior first molar tooth in sambar deer. The infraorbital foramina were small, elliptical and placed at the level of the superior first premolar tooth. The alveolus for a canine tooth was present rostrally in the maxilla of both species. Turbinates bones were visible and mandibular symphysis remained unossified on radiographs and CT in both species. The radiographs of both species showed that the nasal canal was divided by the nasal septum. The CT scan demonstrated the paranasal, frontal and maxillary sinuses. CONCLUSIONS: The present study is important in the comparative anatomy of ruminant species and may help the wildlife forensic officials to identify and differentiate the bones of these two species from those of other domestic and wild small ruminants.

Seasonal distribution and habitat use preference of Barking deer (Muntiacus vaginalis) in Murree-Kotli Sattian-Kahuta National Park, Punjab Pakistan / Distribuição sazonal e preferência de uso de habitat de veados-latidos (Muntiacus vaginalis) no Parque Nacional Murree-Kotli Sattian-Kahuta, Punjab Paquistão

Microhabitat factors associated with the habitat of barking deer (Muntiacus vaginalis) were examined and compared between summer and winter seasons. Habitat characteristics and preferred habitat were measured by locating direct and indirect signs. To quantify the habitat utilization of barking deer, each selected study site was sampled for floral diversity from 2015 to 2017. Quadrats were deployed along transect lines to determine seasonal distribution. Barking deer were not evenly distributed across vegetation types in the study area; they occurred more often in the broad-leaved forest than in Chir pine forest, at an elevational range of 550-850 m, in thick vegetation on steep slopes. The most preferred habitat included trees and shrubs with 30% and 69% cover, respectively. Barking deer avoided thicker tree cover, possibly as it hinders movement and escape from predators. No significant difference (χ2 = 6.37, df = 3, p = 0.19) in seasonal vegetation cover was recorded.

Fatores de micro-hábitat associados ao hábitat do veado (Muntiacus vaginalis) foram examinados e comparados entre as estações de verão e inverno. As características do hábitat e o hábitat preferido foram medidos, localizando sinais diretos e indiretos. Para quantificar a utilização do hábitat de cervos-latidos, cada local de estudo selecionado foi amostrado para a diversidade floral de 2015 a 2017. Quadrats foram implantados ao longo de linhas de transecto para determinar a distribuição sazonal., Veados-latidos não foram distribuídos uniformemente pelos tipos de vegetação na área de estudo; ocorreram com mais frequência na floresta de folhas largas do que na floresta de pinheiros Chir, em uma faixa de elevação de 550-850 m, em vegetação densa, em encostas íngremes. O hábitat mais preferido incluía árvores e arbustos com 30% e 69% de cobertura, respectivamente. O veado-latido evitou uma cobertura de árvores mais espessa, possivelmente porque impede o movimento e a fuga de predadores. Nenhuma diferença significativa (χ2 = 6,37; df = 3; p = 0,19) na cobertura vegetal sazonal foi registrada.

Seasonal distribution and habitat use preference of Barking deer (Muntiacus vaginalis) in Murree-Kotli Sattian-Kahuta National Park, Punjab Pakistan / Distribuição sazonal e preferência de uso de habitat de veados-latidos (Muntiacus vaginalis) no Parque Nacional Murree-Kotli Sattian-Kahuta, Punjab Paquistão

Microhabitat factors associated with the habitat of barking deer (Muntiacus vaginalis) were examined and compared between summer and winter seasons. Habitat characteristics and preferred habitat were measured by locating direct and indirect signs. To quantify the habitat utilization of barking deer, each selected study site was sampled for floral diversity from 2015 to 2017. Quadrats were deployed along transect lines to determine seasonal distribution. Barking deer were not evenly distributed across vegetation types in the study area; they occurred more often in the broad-leaved forest than in Chir pine forest, at an elevational range of 550-850 m, in thick vegetation on steep slopes. The most preferred habitat included trees and shrubs with 30% and 69% cover, respectively. Barking deer avoided thicker tree cover, possibly as it hinders movement and escape from predators. No significant difference (χ2 = 6.37, df = 3, p = 0.19) in seasonal vegetation cover was recorded.

Fatores de micro-hábitat associados ao hábitat do veado (Muntiacus vaginalis) foram examinados e comparados entre as estações de verão e inverno. As características do hábitat e o hábitat preferido foram medidos, localizando sinais diretos e indiretos. Para quantificar a utilização do hábitat de cervos-latidos, cada local de estudo selecionado foi amostrado para a diversidade floral de 2015 a 2017. Quadrats foram implantados ao longo de linhas de transecto para determinar a distribuição sazonal., Veados-latidos não foram distribuídos uniformemente pelos tipos de vegetação na área de estudo; ocorreram com mais frequência na floresta de folhas largas do que na floresta de pinheiros Chir, em uma faixa de elevação de 550-850 m, em vegetação densa, em encostas íngremes. O hábitat mais preferido incluía árvores e arbustos com 30% e 69% de cobertura, respectivamente. O veado-latido evitou uma cobertura de árvores mais espessa, possivelmente porque impede o movimento e a fuga de predadores. Nenhuma diferença significativa (χ2 = 6,37; df = 3; p = 0,19) na cobertura vegetal sazonal foi registrada.

Molecular mechanisms and topological consequences of drastic chromosomal rearrangements of muntjac deer.

Muntjac deer have experienced drastic karyotype changes during their speciation, making it an ideal model for studying mechanisms and functional consequences of mammalian chromosome evolution. Here we generated chromosome-level genomes for Hydropotes inermis (2n = 70), Muntiacus reevesi (2n = 46), female and male M. crinifrons (2n = 8/9) and a contig-level genome for M. gongshanensis (2n = 8/9). These high-quality genomes combined with Hi-C data allowed us to reveal the evolution of 3D chromatin architectures during mammalian chromosome evolution. We find that the chromosome fusion events of muntjac species did not alter the A/B compartment structure and topologically associated domains near the fusion sites, but new chromatin interactions were gradually established across the fusion sites. The recently borne neo-Y chromosome of M. crinifrons, which underwent male-specific inversions, has dramatically restructured chromatin compartments, recapitulating the early evolution of canonical mammalian Y chromosomes. We also reveal that a complex structure containing unique centromeric satellite, truncated telomeric and palindrome repeats might have mediated muntjacs' recurrent chromosome fusions. These results provide insights into the recurrent chromosome tandem fusion in muntjacs, early evolution of mammalian sex chromosomes, and reveal how chromosome rearrangements can reshape the 3D chromatin regulatory conformations during species evolution.

Gene Expression Analysis with No Sequence Data: Study on Reeves's Muntjac (Muntiacus reevesi).

Despite scientific progress, the gene sequences for many species not commonly used in research have not yet been analyzed. This makes it difficult to carry out molecular studies on such animals, as the sequence of genes is the basic information used in many techniques. In this study, we attempt to design primers for a real-time PCR analysis, basing on a comparative analysis of selected gene sequences of species related to Reeves's muntjac (Muntiacus reevesi) and by identifying highly conservative regions. Results of PCR products sequencing and their alignment with the GenBank collection show that all selected primers gave products highly similar (> 90%) to the intended target (among compared species), which led us to the conclusion that our primers may be used for further analyses of gene expression.

Genetic similarities and phylogenetic analysis of Muntjac (Muntiacus spp.) by comparing the nucleotide sequence of 16S rRNA and cytochrome B genome.

This study aimed to identify the phylogenetic similarities among the muntjac (Muntiacus spp.). The phylogenetic similarities among seven major muntjac species were studied by comparing the nucleotide sequence of 16s rRNA and cytochrome b genome. Nucleotide sequences, retrieved from NCBI databases were aligned by using DNASTAR software. A phylogenetic tree was created for the selected species of muntjac by using the maximum likelihood method on MEGA7 software. The results of nucleotide sequences (16s rRNA) showed phylogenetic similarities between, the M. truongsonensis and M. rooseveltorum had the highest (99.2%) while the lowest similarities (96.8%) found between M. crinifrons and M. putaoensi. While the results of nucleotide sequences (Cty b) showed the highest similarity (100%) between M. muntjak and M. truongsonensis and the lowest s (91.5%) among M. putaoensis and M. crinifrons. The phylogenetic tree of muntjac species (16s rRNA gene) shows the main two clusters, the one including M. putaoensis, M. truongsonensis, M. rooseveltorum, and M. muntjak, and the second one including M. crinifrons and M. vuquangensis. The M. reevesi exists separately in the phylogenetic tree. The phylogenetic tree of muntjac species using cytochrome b genes shows that the M. muntjak and M. truongsonensis are clustered in the same group.

Seasonal distribution and habitat use preference of Barking deer (Muntiacus vaginalis) in Murree-Kotli Sattian-Kahuta National Park, Punjab Pakistan.

Microhabitat factors associated with the habitat of barking deer (Muntiacus vaginalis) were examined and compared between summer and winter seasons. Habitat characteristics and preferred habitat were measured by locating direct and indirect signs. To quantify the habitat utilization of barking deer, each selected study site was sampled for floral diversity from 2015 to 2017. Quadrats were deployed along transect lines to determine seasonal distribution. Barking deer were not evenly distributed across vegetation types in the study area; they occurred more often in the broad-leaved forest than in Chir pine forest, at an elevational range of 550-850 m, in thick vegetation on steep slopes. The most preferred habitat included trees and shrubs with 30% and 69% cover, respectively. Barking deer avoided thicker tree cover, possibly as it hinders movement and escape from predators. No significant difference (χ2 = 6.37, df = 3, p = 0.19) in seasonal vegetation cover was recorded.

Phylogeography and population genetic structure of red muntjacs: evidence of enigmatic Himalayan red muntjac from India.

BACKGROUND: Identifying factors shaping population genetic structure across continuous landscapes in the context of biogeographic boundaries for lineage diversification has been a challenging goal. The red muntjacs cover a wide range across multiple vegetation types, making the group an excellent model to study South and Southeast Asian biogeography. Therefore, we analysed mitogenomes and microsatellite loci, confirming the number of red muntjac lineages from India, gaining insights into the evolutionary history and phylogeography of red muntjacs. RESULTS: Our results indicated the Northwestern population of red muntjac or the Himalayan red muntjac (M. aureus) in India as genetically diverse and well-structured, with significant genetic differentiation implying a low level of gene flow. The phylogenetic, population genetic structure, as well as species delimitation analyses, confirm the presence of the lineage from Western Himalayan in addition to the previously identified red muntjac lineages. Relatively low genetic diversity was observed in M. aureus compared to M. vaginalis, M. malabaricus and M. muntjak. The M. aureus and M. vaginalis lineages have split during the late Pleistocene, ~ 1.01 million years ago (Mya), making M. aureus the youngest lineage; whereas, M. malabaricus split earlier, ~ 2.2 Mya and appeared as the oldest lineage among red muntjacs. CONCLUSIONS: Pronounced climate fluctuations during the Quaternary period were pivotal in influencing the current spatial distribution of forest-dwelling species' restriction to Northwestern India. Our finding confirms the distinct Himalayan red muntjac (M. aureus) within the red muntjac group from Northwestern India that should be managed as an Evolutionary Significant Unit (ESU). We recommend a reassessment of the conservation status of red muntjacs for effective conservation and management.

Comparative morphometrical studies on the skull bones of barking deer (Muntiacus muntjak) and sambar deer (Rusa unicolor).

The present study reports data on the skull bone morphometry of barking and sambar deer. The skulls of adult barking deer (n = 6) and sambar deer (n = 6) of either sex (n = 3 males and n = 3 females) were collected from the Aizawl Zoological Park, Aizawl, Mizoram, India, with official permission from the Government of Mizoram. Anatomically, barking and sambar deer's skulls were elongated, pyramid-like, dolichocephalic and consisted of thirty-two cranial and facial bones. The cranial bones were eleven (three single and four paired), comprising of occipital, sphenoid, ethmoid, frontal, interparietal, parietal and temporal. The facial bones were twenty-one (one single and ten were paired), consisting of the maxilla, premaxilla (incisive), palatine, pterygoid, nasal, lacrimal, zygomatic (malar), vomer, turbinates, mandible and hyoid. In the present study, altogether 41 different measurements were taken morphologically and 6 different indices were applied. The obtained morphometrical parameters were significantly (p < .01, p < .05) higher in males than females of both species. Species wise, all obtained parameters were higher in sambar deer than barking deer. The obtained 41 different skull parameters and 6 indices showed statistically significant differences (p < .01 and p < .05) between both sexes of barking and sambar deer; however, practically these differences were meagre. The present morphometrical study on the skull of both species can help the wildlife professionals and zoo veterinarians determine the sex of these animals and differentiate it from other domestic and wild small ruminants for solving veterolegal cases. This study's findings will also motivate and assist other comparative studies with various domestic and wild small ruminants.

Feeding habits and habitat use of barking deer (Muntiacus vaginalis) in Himalayan foothills, Pakistan.

Northern red muntjac (Muntiacus vaginalis; "barking deer") is a shy and small-sized cervid mammal, limited to the outer Himalayan foothill forests in Pakistan. Habitat characteristics were measured by locating direct and indirect signs. To quantify habitat utilization of barking deer, 80 field surveys were conducted in the study area along transects. 1200 Quadrats at 50 m intervals were deployed along these transect lines to determine microhabitat factors associated with seasonal distribution. The food composition of the barking deer was determined through fecal droppings analysis by micro-histological technique. Forty-five fecal samples of barking deer were collected from the study area (Murree-Kotli Sattian-Kahuta National Pak); summer (28) and winter (17). The micro-histological analysis revealed that more plant species are available in its habitat during the summer season (27) as compared to winter (19). Due to browsing nature barking deer mostly feed on trees in both seasons. While shrubs are slightly higher in winters. In summer barking deer consumed 10 Trees, 6 Shrubs, 5 Herbs, and 6 kinds of grass species. Dominant tree species were Phyllanthus emblica and Acacia modesta. Dominant shrub species were Ziziphus nummularia and Justicia adhatoda. In winter barking deer consumed 8 Trees, 7 Shrubs, 3 Herbs, and 1 Grass. Dominant tree species were Bauhinia variegata and Acacia modesta while shrubs included Ziziphus nummularia and Carissa opaca.