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 andstructure 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,
uvFigs.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: 1400xFigs. 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. The findings have made the morphology of
the intestine in the Java treeshrew more closely resembles to
those of the carnivores or omnivores than those of the true
insectivores.ACKNOWLEDGEMENT
‘The authors thank the Directorate General of Nature
Protection and Conservation, Ministry of Forestry and
Plantation Indonesia for permitting the use of the animals in
the present study. Thanks are also due to Dr. Dwi Kesuma
Sari for excellent illustrations. This work was partly funded
by URGE research grant (no. O07/PDB/URGE/1996) from
the Ministry of Edu-cation and Culture, Indonesia.
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