MORPHOLOGICAL STUDY OF THE INTESTINE OF THE JAVA TREESHREW (Tupaia javanica) STUDI MORFOLOGI USUS TUPAIJAWA (Tupaia javanica) ‘Srihadi Agungpriyono', Dwi Liliek Kusindarto’, Chairun Nisa’, Eiichi Hondo’, Masamichi Kurohmaru‘, Junzo Yamada’ Department of Veterinary Anatomy, 'Faculy of Veterinary Medicine, Bogor Agricultural University, J. Taman Kencana 3, Bogor 16151 INDONESIA, Fax. (62-251) 343153, email: ysrihadi@yahoo.com, Faculty of Veterinary Medicine, Gajah Mada University, Vogikarta, INDONESIA, ‘Obihiro University of Agriculture and Veterinary Medicine, Obihiro 080-8555, JAPAN, ‘Faculty of Agriculture, ‘The University of Tokyo, Tokyo, JAPAN ABSTRACT Media Veteriner. 1999. 6(3): 15-21 ‘The morphology of the intestinal tract of Java tree- shrew (Tupaia javanica) was studied using macroscopic, light. and scanning. electron-microscopic methods. The intestine was short in proportion to body length. The proportion of 1:3 was between the insectivores and car- nivores. ‘The large intestine was simple with small caecum ‘and short descending colon, There were no ascending or transverse colon. The mucosal surface of the small intestine showed mucosal folds and intestinal villi, which varied in their shape, size and density depend on the portions of the intestine. The large intestine had longitudinal mucosal folds. ‘The villi were finger- or tongue-like in the duodenal and Jejunal parts, and low thick column-shaped in the ileal part ‘The epithelium and intestinal glands consisted of absorptive cells, goblet cells, and endocrine cells. Eosinophilic granular (Paneth) cells were found in the intestinal glands of small intestine and caecum. The morphology of the intestine of the Java treeshrew is more closely resembles. those of the carni- vores oF omnivores than those of the true insectivores. Key words: alimentary system, histology, intestinal villi, Scandentia ABSTRAK Media Veteriner. 1999, 6(3): 15-21 Detail morfologi anatomi usus tupai Jawa (Tupaia Javanica), dipelajari pada tingkat makroskopis, mikroskopis dan mikroskop elektron scanning. Proporsi usus tupai Jawa relatif pendek terhadap panjang badannya, Panjang badan berbanding panjang usus adalah 1:3. Nilai perbandingan ini bberada diantara hewan insektivora muri dan karnivora. Usus besar menunjukkan susunan yang sederhana dengan sekum yang sangat kecil dan kolon descenden yang pendek. Tidak dijumpai adanya kolon ascenden dan kolon trans- versum, Mukosa usus Kecil ditandai oleh adanya vili usus dengan bentuk-bentuk seperti jarum, jari atau lidah pada duodenum dan yeyunum, serta bentuk seperti Kolom tebal ‘yang rendah pada ileum. Pada usus besar tidak dijumpai vi iecuali lipatan mukosa yang berjalan longitudinal. Sel-sel pada lapis epitel dan Kelenjar usus terdiri atas sel-sel Penyerap, sel goblet dan sel endokrin, Selain itu dijumpai pula sel Pancth pada kelenjar usus di usus kecil dan sekum. Gambaran morfologiusus tupai Jawa lebih banyak me- nycrupai gambaran morfologi pada hewan karnivora atau ‘omnivora dibandingkan dengan hewan insektivora murni Katackata kunci: sistem pencernaan, histologi, vill usus, Scandentia INTRODUCTION The Java treeshrew (Tupaia javanica) is one species of the treeshrews that commonly found in Java Island. The nimal is insectivorous or omnivorous, has mixed dict of insects, worms, fruits, seeds and buds (Lyon, 1913). Simple stomach and short intestine with or without small caecum ‘characterize the alimentary tract of this animal (Lyon, 1913; Luckett, 1980; Makita et al., 1998; Agungpriyono et al, 1999). ‘The morphophysiology of the gastrointestinal tract of the Java treeshrew has not been clearly understood as only partial studies available. These studies include gross anato- mical reports by Lyon (1913) on the gastrointestinal tract and Makita ef al. (1998) on the caecum and a report by ‘Agungpriyono et al. (1999) on the stomach morphology. To provide a beiter understanding about the digestive system of this species, morphological characteristics of the intestinal tract such as surface structure of intestinal villi andstructure of the lining intestinal epithelial and glandular cells should be clarified. In this study, the morphological characteristics of the intestinal tract of the Java treeshrew were described using macroscopic, scanning electron- and light- microscopic methods. MATERIAL AND METHODS Ten animals of T: javanica of both sexes, weighing 70- 120 g, were used in this study. The animals were catched under license from their habitat in the eastern and central parts of Java Island. After a stabilization period of one week, the animals were sacrificed by bleeding after a deep anesthesia using pentobarbitone (Nembutal®, Abbot Lab., Miinois, USA). Afier observation of in situ position, whole digestive tract was removed out from the body, washed in 0.1 M phosphate buffered saline (PBS, pH 7.4) and fixed in Bouin fluid for 24 hours. To facilitate better penetration of fixative solution, the fixative solution was also injected into the Iu- ‘men. Macroscopic observations were made with naked eye or with a stereo dissecting microscope (Zeiss, Germany) Scanning Electron Microscopy ‘Tissue samples from duodenum to rectum were taken and processed for scanning electron microscopy. Tissues were washed well with PBS solution and immersed for 2 ours in 2% tannic acid. This was followed by post-fixation in 1% osmium tetroxide for 1 hour at room temperature (Murakami, 1973). All samples were then dehydrated using. ‘butyl alcohol freéze drying method (Inoue and Osatake, 1988), mounted on the metal stubs, coated with gold by sputtering and observed under a scanning electron micro- scope (JEOL, JSM-35CF, Japan) at an accelerating voltage 10kV. Light Microscopy Tissues from certain portions as above were processed for embedding in paraffin. These included dehyération of the tissues in a graded series of ethanol-xylol and embedding in paraffin. Paraffin sections were cut serially at 5 um of thickness. Sections were deparaffinized, rehydrated and then stained with hemstoxylin-eosin for observation of general structure and with Grimelius silver impregnation staining ‘method (Grimelius, 1968) for observation of endocrine cells. Sections were observed and photographed using a Tight microscope (Olympus Vanox, Japan). RESULTS AND DISCUSSION ‘Based on our observations and measurement, the length of the intestine of the Java treeshrew (T. javanica), was relatively short in its proportion to the body length, 1:3 (able 1). Both small and large intestinal parts were pre sent. Duodenal and rectal parts of the intestine were identified obviously by their location next to the pylorus and anus, respectively. Large intestine was simple, The caecum ‘was markedly small, and there were no ascending or trans- verse colon, except a short descending colon. The Java tree- shrew was noted to have short intestine as compared to their body length, 1:3. This proportion stands between the insectivores (1:2) and the carnivores (1:4) (see Table 2). Generally, the intestine of carnivores, especially the insecti- vvorous animals, are considerably shorter than those of her- bivores, and those of omnivores are intermediate in length between the carnivores and herbivores (Nickel et al. 1973; Kurohmaru et al., 1980, Steven, 1988). The proportion in the Java treeshrew is coincide with the fact that the animal diets not only on insect and worms but also on fruits, seeds and buds (Lyon, 1913). The caecum of the insectivorous animals is very small or absent (Kurohmart et al.. 1980). Tnsectivorous animals, musk shrew and long-tailed shrew, are reported to have no caecum (Kurohmaru et al., 1980; Kurohmaru et al., 1982). The caecum in the Java treeshrew ‘was very small. This finding reconfirms the previous report of Makita et al. (1998). Based on above findings itis sug ‘gested that the Java treeshrew may have more carnivorous or omnivorous nature than the true insectivores such as musk shrew or long-tailed shrew. ‘The mucosal surface of the smait intestine showed mu- cosal folds and intestinal vii. The intestinal villi (Figs. 1, 2) varied in their shape, size and density depend on the portions of the intestine. The density of the villi was high in the proximal intestine, ie. the duodenum. The shape of each villus of the intestine was leaf- or tongue-like in the duodenal and jejunal parts, and low thick column-shaped or ridge-like in the ileal part. The height of the villi was decreased towards the caudal portion. White the small in- testine showed intestinal villi, the large intestine possessed longitudinal mucosal folds running parallel from colon to rectum, Circular folds (Plica circulares) was not developed ‘Table 1. Mean length (cm) of the gastrointestinal portions of the Java treeshrew Length of Distance between orifices Length of small _Lengih of Length of large Stomach esophagus to duodenum intestine (duodenum to Caecum —_intestine (colon to ileum) rectum) 42 032, 375 098 6.46 16' Table 2. Rough ratios of the body length to the intestine length (excluding tail) “Animal species ‘Type of animal ‘Reference tailed shrew Taeativorous ‘Kurohmaru et al. (1980) Musk shrew Insectivorous ‘Kurohmaru et al. (1980) Java treeshrew Inseetivorous/ omnivorous ‘Present study Cat Carnivorous Nickel et al. (1973) Dog Carnivorous Nickel etal. (1973) Hamster, rat Ornivorous Kurohmaru eral. (1980) Mouse Ornnivorous Nickel etal. (1973) Human Omnivorous Nickel etal. (1973) Horse Herbivorous ‘Nickel eal. (1973) Pig Omnivorous Nickel etal. (1973) Ox Herbivorous Nickel et al. (1973) Sheep-Goat Herbivorous Nickel etal. (1973) villous morphology showed variations as those of musk shrew (Kurohmaru ef al., 1980), oot findings on the distribution of intestinal villi do not coincide with that of rusk shrew. In musk shrew, intestinal villi are distributed up ‘othe large intestine. ‘The intestinal epithelium (Figs. 3A,C) consisted of ab- sorptive epithelial cells, mucous secreting goblet cells and basal granulated cells or endocrine cells. Each absorptive epithelial cell had an oval nucleus in the lower part of me cell body and well developed microvilli on its free apical surface (Fig. 3C). Many plasma cells and lymphocytes were observed in the core of each intestinal villus. Capillary blood ‘yescels were present in the thick submucosal part. Duodenal Brunner) glands (Fig. 3D) were observed to a limited Fer extends about 1 cm from the pylorus. Lymphatic nodules Fig. 1. Mlustation ofthe gastrointestinal tract ofthe Java treeshrew (body length : 15 em) inall parts of the intestine, except in the rectal part. ‘The morphology of the intestinal villi is stated to vary depend on their location in the small intestine (Woodal and Skinner, 1994). Villi in the proximai small intestine are longer and numerous, which may represent a high degree of digestive process such as absorption and secretion. The form of the villi varies between aninal species (Taylor and Anderson, 1972; Dellmann, 1993) and is correlated well with animal diet (Woodal and Skinner 1994). We observed similar condition in the intestinal vil ofthe Java tree-shrew. ‘The villi were leaf- or tongue-like shaped in the duodenal and jejunal parts, and low thick column-shaped or ridge-like in the ileal part. The villi were decreased in their height towards caudal portion. Although our present result on the were often observed in the transitional areas. The present findings on the histological structure of lining epithelial cells resemble those of other mammals. Absorptive epithelial cells had microvilli on their free apical border. The microvilli are formed to enlarge the surface for absorption process (Dellmann, 1993). Goblet cells were observed along the intestinal tract and their number was numerous in the large intestine (Fig. 44). Under scanning electron microscopy, the secreted mucous appeared as large bleb on the apical portion of the goblet cells (Figs. 2C,D). Mucous secreting goblet cells were more ‘numerous in the large intestine. Similar condition is reported for other mammals (Trautmann and Fiebiger, 19575 Dellmann, 1993; Ross et al., 1995). The mucous is needed for mucosal protection and smooth passing of the ingesta through the large intestine. Intestinal gland was simple tubular consisted of similar cells as the epithelium (Fig. 3B). In addition ta these cells, uvFigs.2. Scanning electron micrographs of the intestinal mucos cosal surface of large intestine (D). Intestinal villi are tongue- or finger-like in duodenum (A) and leaf- or ridge-Hi tum (B). The epithelial cells are clearly seen at higher mag Mucous secreted by the goblet cells are seen on the surface A,B.D: 100x; C : 400x ik on (C), being round, oval, pyramid or polyhedral shapes, 1). Tke mucosa of colon (large intestine) has no villi (D),Figs. 3. Light photomicrographs of the intestinal mucosa. The lining epithelial cells (A, C) consist of columnar absorptive cells (1) and mucous secreting goblet vells (2). Intestinal glands or glands of Licberkuhn (B) are located below the epithelium, consist of the same cells a5 the epithelial layer. Duodenal (Brunner) glands (D) are found in the submucosa of duodenum, consist of basophilic mucous cells. Eosinophilic granular (Paneth) cells (E, arrows) are found in the basal portion of the intestinal glands. Hematoxylin-eosin stain. A, B,D : 5S0x; C, E: 1400xFigs. 4. Light photomicrographs of the large (A) and small (B) intestines showing goblet cells (A, arrows) and endocrine cells (B, arrows) in the intestinal mucosa, Goblet cells are numerous inthe large intestine. Gut endocrine cells are stained dark brown with Grimelius silver impregnation method. ‘The cells are round, oval or triangular, mainly located in the ‘basal portions ofthe intestinal glands. A : Hematoxylin-eosin stain. B : Grimelius stain. A.B :275x. cosinophilic granular (Paneth) cells were observed in the intestinal glands of the proximal portion of the intestine, ie. from duodenum to caecum, Under light microscopy, these cells showed characteristic features by their basal location in the intestinal glands and by having bright eosinophilic ‘granules in their cytoplasm (Fig. 3E). Paneth cells are function in the production of peroxidase and antibacterial ‘compound called lysozyme (Dellmann and Carithers, 1996). ‘The presence of Paneth celis in the intestinal gland of the Java treeshrew is similar with human, thesus monkey. hare, guinea pig, rat, mouse, golden hamster, insectivores bats (Satoh ef al, 1990; Dellmann, 1993), ruminant and horses (Dellmann, 1993; Dellmany and Carithers, 1996). The Paneth cells are absent in carnivores, pig (Dellmann and Carithers, 1996), musk shrew (Kurohmaru et al., 1980) and Jong-tailed shrew (Kurohmaru et al, 1982). Endocrine cells (Fig. 4B) were round, oval or triangular and characterized by the presence of cytoplasmic granules at their basal portion. The endocrine cells were observed in all portions examined. They were more numerous in the terminal portions such as duodenum, ileum and rectum. The endocrine cells secrete hormones for regulation of digestive process. The cells are dispersed among the cells of 20 epithelium and intestinal glands along the intestinal tract. ‘The morphotogical characteristic of the endocrine cells is similar with those reported for other mammals (Polak, 1989). Detail data on kinds and distribution of gut endo- crine cells in the intestinal tract of the Java treeshrew has bbeen published separately (Agungpriyono et al., 1998). CONCLUSION ‘Some characteristics on the morphology of the intestine of the Java teeshrew were noted in this study. These ii clude the ratio of body length to the intestinal length, simple structure of the large intestine with small caecum and short colon, the distribution of intestinal villi only in the small intestine, the composition of cells in the epithelium and intestinal glands including the presence of Paneth cells in the intestinal glands. 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