Kanchil Research Articles

Reproductive characteristics of female lesser mouse deers (Tragulus javanicus) based on fecal progestagens and breeding records

Information about breeding and the reproductive biology of mouse deer is limited in the wild and captivity. No reports on reproductive endocrinology are available. The objective of the present study was to observe the reproductive biology based on breeding records, to validate the utility of the non-invasive endocrine monitoring technique using feces of the female lesser mouse deer (Tragulus javanicus), and thus to clarify the reproductive physiology. Breeding records from 2 females were investigated and the fecal progestagen profile was monitored in captivity. Fecal progestagens were extracted using methanol and measured by enzyme immunoassay. From the breeding records, many births occurred in May (spring) and November–December (winter); however, fecal progestagen profile showed cyclical changes throughout the year in a female mouse deer. Most mounting and mating behaviors were observed 2–3 days after the peak of progestagen concentration during luteal phase. The ovarian cycle length based on the fecal progestagen profile averaged 14.5±0.3 days. The fecal progestagen concentration remained high during pregnancy. Fecal progestagen monitoring is useful for evaluating ovarian activity and pregnancy in the lesser mouse deer.

Effects of diets of differing fiber contents on digestibility, passage rate of digesta and heat production in lesser mouse deer (Tragulus javanicus)

The effects of different levels of dietary fiber on feed intake, digestibility, mean retention times of solute and particle and heat production were studied in twelve male lesser mouse deer (Tragulus javanicus). The animals were randomly assigned into four groups of three mouse deer each and fed on individual basis. Four diets, namely A, B, C and D were prepared in pellet forms. Each diet contained ∼14% crude protein and ∼18kJ/g gross energy. The crude fiber contents of diet A, B, C and D were 4.2, 11.3, 16.8 and 22.5%, respectively. The results showed that the crude fiber intake of mouse deer was 1.4±0.04, 3.8±0.4, 5.8±0.7 and 6.0±0.6g/kg W0.75/d fed diet A, B, C and D, respectively. Mouse deer fed diet D had significantly lower DM intake than those fed lower levels of fiber. The digestibility values of DM decreased gradually with increasing levels of fiber in the diet. The mean retention times (MRTs) of particles (∼1.5mm) in the reticulorumen were in the range of (19.0–22.9h), with the shortest time for mouse deer fed diet D. However mouse deer fed diet C and D showed significant shorter solute MRT in the reticulorumen (17.1–18.5h) when compared to mouse deer fed lower fiber diets (21.4–21.9h). The selectivity factor in the reticulorumen was in the range of 1.04–1.18, indicating the mouse deer to be a ‘moose-type’ ruminant. Fiber levels had no significant effect on water intake of mouse deer. Heat production tended to decrease with increasing levels of fiber (448.3–435.7kJ/kgW0.75/d) but differences among the mouse deer fed the four diets were not significant.

Anatomical and histological characterization of ileal and jejunal Peyer's patch in lesser mouse deer (Tragulus javanicus)

Tragulidae is a primitive ungulate family within the order Cetartiodactyla, suborder Ruminantia. Domestic ruminants such as cattle, sheep, and goat have two types of Peyer's patches (PP): jejunal and ileal PP, in which there are morphological and functional differences. In this study, lesser mouse deer (Tragulus javanicus) PP was studied by gross anatomical and histological procedures. At the fetal stage, both types of PP were formed in the small intestine. Ileal PP was observed as a single continuous aggregation of lymphatic follicles extending cranially from the ileo-caecal junction. However, jejunal PP was observed as multiple and discrete accumulations of lymphatic follicles. This study showed that the lesser mouse deer has two types of PP in the small intestine. In addition, the anatomical and histological characteristics of jejunal and ileal PP are quite similar to those of other ruminants’ jejunal and ileal PP. Further studies are needed to analyze immune function of both PP in lesser mouse deer in order to determine the evolutionary process of Cetartiodactyla.

Energy requirement for maintenance of growing and adult male lesser mouse deer ( Tragulus javanicus) in captivity

A study was conducted to determine the energy requirement for maintenance of eight lesser mouse deer, consisting of four growing males (6–8 months of age) with average body weight of 1.4 ± 0.2 kg and four adult males (>2 years of age) with average body weight of 1.6 ± 0.2 kg. The animals were from the colony reared at the Department of Animal Science, Universiti Putra Malaysia. The experiment was based on a 2 × 4 factorial arrangement in Latin-Square design. The animals were kept in single pens and fed with different levels of feed (70%, 80%, 90% and 100% of ad libitum intake). The feed was made up of 95% pellet and 5% ‘kangkong’ ( Ipomoea aquatica) leaves. The energy requirement for maintenance was determined by using the regression equations between metabolisable energy intake and energy retention or body weight change. Energy retention was calculated as the difference between metabolisable energy intake and heat production. The results showed that the metabolisable energy requirements for maintenance when calculated based on energy retention were 420.1 and 422.9 kJ kg −0.75 d −1 for growing and adult mouse deer, respectively, and when calculated based on weight change the values were 435.1 and 436.9 kJ kg −0.75 d −1, respectively. The efficiency of utilization of metabolisable energy for maintenance ( k m ) for growing mouse deer was 0.58 and that for adult mouse deer was 0.54.

Ectoparasites of lesser mouse eared bat, Myotis blythii from Kermanshah Iran

Objective To identify the ectoparasits of lesser mouse eared bat, Myotis blythii inside the Mahi-Dasth cave in Kermanshah province, Iran. Methods A total of 30 lasser mouse eared bat ( Myotis blythii) in a cave close to Kermanshah city were hunted with special net and were transported to the laboratory. After anesthetizing by chloroform, ectoparasites from these bats were removed and identified. Results The ectoparasites include flies, ticks and mites. The 39 identified flies belonged to two families including Strebilidae (7.7%) and Nycteribiidae (92.3%). A total number of 5 ticks of larval, nymphal and adult stages of Ixodes vespertilionis and 18 mites from Spiturnicidae family were identified. Conclusions The result of this study is a clue for the identification of ectoparasites from bats and implication of possible prevention measures for diseases transmitted by ectoparasites.

Immunohistochemical Localization of the Cytoskeletal Proteins in the Testes of the Lesser Mouse Deer (Tragulus javanicus)

Abstract. This study demonstrated the immunohistochemical localization of cytoskeletal proteins, cytokeratin, desmin, vimentin, α-tubulin and α-smooth muscle actin (α-SMA) in the testes of adult and immature lesser mouse deer (Tragulus javanicus). Desmin was found in the sub-epithelial myoid cells of epididymal ducts. The immunoreactivity for vimentin was shown in the peritubular myoid cells and around the nuclei of the Sertoli cells of which the positions indicated various arrangements. In the Sertoli cells, vimentin filaments between the basement membrane and the nucleus were elongated with the migration of the nucleus toward the upper part. Furthermore, vimentin was detected in the Leydig cells and the sub-epithelial myoid cells of epididymal ducts. Alpha-tubulin was apparently extended to the tip of each Sertoli cell cytoplasm, and α-SMA was found in the peritubular myoid cells and the sub-epithelial myoid cells of epididymal ducts. On the other hand, cytokeratin and desmin were not detected in the Se...

Morfologi dan Biometri Spermatozoa Anoa (Bubalus Sp.) yang Diwarnai dengan Pewarna William’s dan Eosin-Nigrosin

Lesser mouse deer is a native animal, which is commonly found in the forest of Java and information concerning this animal is still limited. This research explored the quality of lesser mouse deer meat ( Tragulus javanicus ). Eight lesser mouse deer (four females and four males) were used in quality aspects of lesser mouse deer meat. The physical properties observed were pH, water holding capacity, tenderness, and cooking loss. While the chemical properties observed were proximate analysis, amino acids, fatty acids, cholesterol, and EPA-DHA of the meat. The results showed that meat samples had average of ultimate pH (6.32), water holding capacity (32.82%), tenderness (1.80 kg f/m2), and cooking loss (45.15%). The average meat chemical compositions namely content of water, protein, fat, ash and cholesterol were 76.33%, 21.42%, 0.51%, 1.20%, and 50.00 mg/100 g, respectively. Fatty acids consisted of lauric, miristic, palmitic, stearic, oleic, linoleic, and linolenic acid were in the amount of 1.04% 3.09%, 30.97%, 0.77%, 59.41%, 3.22%, and 1.12%, respectively. The total EPA and DHA was 0.13% and 0.05%. Respectively, the results showed that quality aspects of lesser mouse deer meat had a high ultimate pH, low water holding capacity, tenderness, high cooking loss, and high water content, high protein, low fat and high cholesterol.

Distribution of Lectin‐Bindings in the Testis of the Lesser Mouse Deer, Tragulus javanicus

The distribution of lectin bindings in the testis of the smallest ruminant, lesser mouse deer (Tragulus javanicus), was studied using 12 biotinylated lectins specific for d-galactose (peanut agglutinin PNA, Ricinus communis agglutinin RCA I), N-acetyl-d-galactosamine (Dolichos biflorus agglutinin DBA, Vicia villosa agglutinin VVA, Soybean agglutinin SBA), N-acetyl-D-glucosamine and sialic acid (wheat germ agglutinin WGA, s-WGA), D-mannose and d-glucose (Lens culinaris agglutinin LCA, Pisum sativum agglutinin PSA, Concanavalin A Con A), L-fucose (Ulex europaeus agglutinin UEA I), and oligosaccharide (Phaseolus vulgaris agglutinin PHA-E) sugar residues. In Golgi-, cap-, and acrosome-phase spermatids, lectin-bindings were found in the acrosome (PNA, RCA I, VVA, SBA, WGA and s-WGA), and in the cytoplasm (PNA, RCA I, VVA, SBA, WGA, LCA, PSA, Con A and PHA-E). s-WGA binding was confined to the spermatid acrosome, but other lectins were also observed in spermatocytes. In spermatogonia, VVA, WGA, Con A, and PHA-E bindings were observed. Sertoli cells were intensely stained with DBA and Con A, and weakly with PHA-E. In interstitial Leydig cells, RCA I, DBA, VVA, Con A, PSA, LCA, WGA and PHA-E were positive. UEA I was negative in all cell types including spermatogenic cells. Unusual distribution of lectin-bindings noted in the testis of lesser mouse deer included the limited distribution of s-WGA only in the spermatid acrosome, the distribution of DBA in Sertoli cells, Leydig cells and lamina propria, and the absence of UEA I in all type cells. The present results were discussed in comparison with those of other animals and their possible functional implications.

Scanning electron microscopic studies on the connective tissue cores of the lingual papillae of lesser mouse deer

Scanning electron microscopic studies on the connective tissue cores of the lingual papillae of lesser mouse deer

Novel Erythrocyte Pits in the Small Tropical Ruminant, Lesser Mouse Deer

We examined unique erythrocyte pits of the peripheral blood and bone marrow in the lesser mouse deer, Tragulus javanicus, using scanning electron microscope (SEM) and transmission electron microscope (TEM). Under the SEM observation, the pit was observed as a hole on both mature erythrocytes of the peripheral blood and immature erythrocytes of the bone marrow. By the TEM, the mature erythrocytes had a vacuole, which showed complicated shape and occupied considerable space within the cytoplasm. The vacuole was communicated extracellularly by perforation, which corresponded to the hole on the cell surface. In the bone marrow, erythroblast and reticulocytes have a cytoplasmic vacuole. This abnormal feature of the erythrocytes is peculiar to the mouse deer, and not found in other tropical ruminants. Despite the disadvantage of volume loss from the small erythrocytes, the mouse deer were healthy and showed no signs of anaemia.

Functional strategies of the hindlimb muscles in the mouse deer

ABSTRACT We observed the muscles of hindlimb, talus, and tarsal joints in the lesser mouse deer (Tragulus javanicus) and greater mouse deer (Tragulus napu) that have conserved the original morphological characters of the musculoskeletal system in the Ruminantia. The proximal trochlea articulating to the distal end of the tibia obviously turned at a medio-proximal direction. Since the proximal parts of the muscle bundles were concentrated in the lateral side of the thigh, the tension of the extensor muscles in the hindlimb is mediodistally conducted. It indicates that the extending power of hindlimb muscles is not parallel with the direction of the trochlea, but crosses the angle of the proximal pulleys in the talus. We suggest that the crossing may enable the animal to stand and run stably and to bear its body weight. We compared the development of the hindlimb muscles in the mouse deer (Tragulus javanicus and Tragulus napu) with that in the Sika deer (Cervus nippon) and Chinese muntjac (Muntiacus reevesi...

Morphology of the Intermandibular Gland of the Lesser Mouse Deer, Tragulus javanicus

The morphology of the intermandibular gland of the Lesser mouse deer (Tragulus javanicus), which plays an important function in marking area and territory and in the reproductive behaviour of the animal, was examined using immunohistochemistry, lectin histochemistry and scanning electron microscopy. The gland was composed of sebaceous and apocrine glandular material. Sebaceous glands occupied a greater area of the total gland and consisted of many large lobules with polyhedral cells having a pale cytoplasm. The sebaceous gland, being holocrine, possessed no special secretory ducts. The apocrine gland was lined by cuboid cells and the secretory products were often seen in the apical portions of the cells. Myoepithelial cells contained actin filaments lining the basal membranes of the apocrine gland and were surrounded by nerve fibres which immunostained with protein gene product 9.5. The secretion of the gland appears to be a mixture of larger amounts of lipid material from sebaceous glands, and glycoconjugates secreted by both sebaceous and apocrine glands. Lectin histochemistry detected these as galactose, N-acetyl-D-galactosamine, N-acetyl-D-glucosamine, D-mannose and D-glucose. The male gland was larger in size and contained more N-acetyl galactosamine and N-acetyl glucosamine in its secretion than the gland of the female. This implied the presence of sexual differences in secretions in the intermandibular gland of the Lesser mouse deer.

A Tandem Gene Duplication Followed by Recruitment of a Retrotransposon Created the Paralogous Bucentaur Gene (bcntp97) in the Ancestral Ruminant

Retrotransposable element-1 (RTE-1) is a class of long interspersed nucleotide elements that contain in its open reading frame an apurinic/apyrimidinic endonuclease domain (AP-END) and a reverse transcriptase domain. Ruminants have a clade-specific RTE-1 (BovB/RTE). The bovine bcnt gene (bucentaur or craniofacial developmental protein 1) has a duplicated paralog (bcntp97) in tandem that recruited an AP-END of BovB/RTE as a coding exon (RTE exon). We obtained sequence of the bcnt region from several animals and showed that other ruminants also have the bcntp97 with a conserved RTE exon while camels and pigs do not. Genomic Southern analysis showed that camels and pigs have multiple bcnt-related sequences but not BovB/RTE which bovines and lesser mouse deer have abundantly. These results indicate that the bcnt gene duplication followed by the creation of bcntp97 including recruitment of the RTE exon occurred in the ancestral ruminant about 55 MYA. The indication of time frame is supported by a phylogenetic analysis. Taken together with a result of differential tissue expression of the two bcnt paralogs, we conclude that bcntp97 was created concurrently with the early radiation of BovB/RTE in an ancestral ruminant and then acquired a novel function.

An Ultrastructural Study on the Leydig and Sertoli Cells in the Immature Lesser Mouse Deer (Tragulus javanicus)

Leydig and Sertoli cells of the immature lesser mouse deer testes, obtained in East Malaysia, were observed using light and transmission electron microscopy (TEM). The testes were fixed in 5% glutaraldehyde, post-fixed in 1% OsO4, dehydrated in ethanol, and embedded in Araldite M. Serial semi-thin sections were cut, stained with toluidine blue and observed using light microscopy. Serial ultra-thin sections were cut, stained with uranyl acetate and lead citrate, and examined using TEM. As a result, ultrastructurally, two types of underdeveloped filament bundles were infrequently recognized in Leydig cells, but not in other testicular cells. One type was the underdeveloped bundles of actin filaments (approximately 5 nm in diameter), which were found in the nucleus of Leydig cells. The other type was the underdeveloped bundles of intermediate filaments (approximately 10 nm in diameter), which were found in the cytoplasm of Leydig cells. A multivesicular nuclear body (MNB)--specifically present in the Sertoli cell nucleus of ruminant testes--was infrequently observed. The MNB is situated in the vicinity of nuclear membrane, still in an underdeveloped stage.

Cytogenetic comparison of the lesser mouse deer (Tragulus javanicus) and the greater mouse deer (T. napu)

Mouse deer is known as the smallest and one of the most primitive ruminants. The lesser and the greater mouse deer originated in three different parts of Malaysia were examined for cytogenetic study. The chromosome number of the mouse deer was 2n=32, consisted of 15 metacentric autosome pairs and X and Y sex chromosome pair. G-, R-band revealed polymorphisms of sex chromosomes among the mouse deer from different origins. X chro- mosome varied in length of short arm between the lesser and the greater mouse deer. Y chromosome showed poly- morphisms not only between the lesser and the greater mouse deer, but also among the lesser mouse deer from dif- ferent origins. The 18S-28S rRNA genes (18S-28S rDNA) were localized only in the X and Y chromosomes in both lesser and greater mouse deer by the Fluorescence in situ hybridization (FISH) method. The hybridization signals of 18S-28S rDNA showed polymorphisms not only in size, but also in their distribution patterns on the sex chromo- somes. In addition to the general banding methods, the FISH analysis determined that the greater mouse deer from Tioman island would be remarkably different from the lesser mouse deer. The cytogenetic observations investigated in this study could be informative to characterize the lesser and the greater mouse deer. And it is also suggested that the chromosomal change would reflect that the relationship between the geographic distribution and speciation among the mouse deer.

Geographical Variation of the Skull of the Lesser Mouse Deer

We examined the geographical variation of the skull size and shape of the lesser mouse deer (Tragulus javanicus) from Laos, Thailand, Peninsular Malaysia, Sumatra, Java, Borneo, Langkawi and some Islands of Tenasserim in Myanmar. Although the influence of the climatic condition on skull size was not confirmed in the mainland populations, the skull became rostro-caudally longer in the populations of Tenasserim and Sumatra because of island isolation effect. The skull size was classified into the following three clusters of localities from the matrix of Q-mode correlation coefficients: 1) Langkawi and Tenasserim, 2) Laos and Thailand, 3) Sumatra and Borneo. The skulls in the population of Java belong to the cluster of Langkawi and Tenasserim in male, however were morphologically similar to those in the cluster of Borneo and Sumatra. The canonical discriminant analysis pointed out that the Laos and Tenasserim populations were separated from the other ones and that the populations of Sumatra, Java and Borneo were intermingled each other.

Phylogenetic relationships among populations of the mouse deer in the Southeast Asian Region from the nucleotide sequence of cytochrome b gene

We examined the phylogeographic structure of the Lesser Mouse deer (Tragulus javanicus) and the Greater Mouse deer (Tragulus napu) from sequence data of mitochondrial cytochrome b gene. Three major clusters were observed on phylogenetic trees, as follows: 1) T. javanicus from the Asian mainland (Malayan Peninsula and Laos), 2) T. javanicus from Borneo Island (Sabah, Malaysia), 3) T. napu from Borneo and Pulau Tioman. Tragulus napu forms the sister lineage to all populations of T. javanicus, thereby supporting the current separation of these taxa at species level and traditional taxonomical identification of the two species by throat color. Within T. javanicus, the primary genetic separation is between the Borneo Island population and those from the Southeast Asian mainland. We suggest that subspecies should be recognised within this widespread taxon. A secondary separation within T. javanicus is observed between populations to the north and south of the Isthmus of Kra.

Peculiar bundles of filaments in Leydig cells of the lesser mouse deer (Tragulus javanicus): an ultrastructural study.

Leydig cells of lesser mouse deer (Tragulus javanicus) testes were observed using light and transmission electron microscopies. Sexually mature lesser mouse deer were obtained in East Malaysia. The testes were perfused with 5% glutaraldehyde, postfixed with 1% OsO4, dehydrated in ethanol and embedded in Araldite. The semithin sections were cut, stained with toluidine blue and observed under light microscopy. The ultrathin sections were cut, stained with uranyl acetate and lead citrate, and examined using a JEM-1200 transmission electron microscope. As a result, two types of filament bundles were frequently recognized in Leydig cells, but not in other testicular cells. These bundles were clearly seen at even a light microscopic level. One type was bundles of actin filaments (approximately 5 nm in diameter). These structures were found not only in the cytoplasm but also in the nucleus. The other type was bundles of intermediate filaments (approximately 10 nm in diameter). These structures were found only in the cytoplasm. The existence of filament bundles has never been reported in the testicular cells of another mammalian species. Thus, while bundles of actin and intermediate filaments are specifically present in the Leydig cells of the lesser mouse deer, their functions are still unclear.

Calpain 3 is expressed in astrocytes of rat and Microcebus brain

The calcium-dependent protease calpain is involved in numerous functions, including the control of cell survival, plasticity and motility. Whereas the isoforms calpain 1 and 2 have been described as ubiquitously expressed enzymes, calpain 3 has been called “muscle-specific”, although trace amounts of calpain 3 mRNA have been detected by Northern blot in brain homogenates. In this study, we validated antibodies raised either against the peptides that were specific for a given isoform or the peptides present in all the three isoforms. We then used the anti-calpain 3 antibodies together with antibodies directed against cell-type-specific proteins to determine by double- and triple-labelling immunocytochemistry if the protease is expressed in specific cell populations of rat as well as lesser mouse lemur ( Microcebus murinus) brain. Calpain 3 was almost exclusively found in cells displaying astrocyte morphology. These cells, most of which co-expressed glial fibrillary acidic protein, were particularly numerous close to the striatal subventricular zone (where numerous neurones forming the rostral migratory stream (RMS) towards the olfactory bulbs are generated) and the RMS itself. Other immunoreactive cells were found close to the pial surface of the forebrain, in the corpus callosum and in the dentate gyrus. Calpain 3 may be involved in astrocyte plasticity and/or motility.

Multivesicular nuclear body in sertoli cells of the lesser mouse deer, Tragulus javanicus.

The Sertoli cell of the lesser mouse deer, Tragulus javanicus, was examined using light and transmission electron microscopy. Similar to other ruminants, a multivesicular nuclear body (MNB) and laminated smooth endoplasmic reticulum (sER) were observed in the lesser mouse deer Sertoli cell. The MNB was present within the Sertoli cell nucleus, and consisted of vesicles, irregular tubules and ribosome-like structures. It was infrequent in the lesser mouse deer, which differs from domestic ruminants. Vesicles and irregular tubules seem to contain some materials with low and/or middle electron density, and be surrounded by electron dense materials. The diameter of vesicles was between 30 nm and 180 nm. Since the MNB, though less developed compared to that of bulls and goats, was present even in the Sertoli cell nucleus of the primitive ruminant-lesser mouse deer, it should be a common structure of ruminant Sertoli cells.