Parasitol Res (2009) 104:701–703

DOI 10.1007/s00436-008-1268-x

SHORT COMMUNICATION

Animal models for Opisthorchis viverrini infection

Thidarut Boonmars & Sirintip Boonjaraspinyo &
Butsara Kaewsamut

Received: 3 November 2008 / Accepted: 5 November 2008 / Published online: 3 December 2008
# Springer-Verlag 2008

Abstract We investigated the utility of various animal infection is considered one of primary causes of cholangio-
models for the study of opisthorchiasis in humans and its carcinoma (CCA) development (Sripa et al. 2007). Studies of
common sequel of cholangiocarcinoma (CCA). Rats, mice, the pathological changes in humans are needed but not
gerbils, and hamsters were infected with Opisthorchis ethically possible so animal models are used.
viverrini metacercariae. Worms from the infected animal Hamster models are the most common animal models of
hosts were recovered from livers and counts made of eggs O. viverrini infection and CCA; despite the time it takes to
per gram of feces. Worms were observed in and recovered establish the infection and subsequent cholangiocarcinoma,
from hamsters and gerbils but not rats and mice. The which will cause the pathological changes to be studied.
recovered worms from the infected gerbils were larger and We, therefore, studied various animal models for their
more physiologically developed than those from the susceptibility to O. viverrini infection and suitability for
infected hamsters. The results suggest that gerbils are more investigation of the pathological changes caused by CCA.
susceptible to infection by Opisthorchis viverrini and thus
more suitable for modeling opisthorchiasis and its connection
to CCA. Material and methods

Parasite preparation
Introduction
O. viverrini metacercariae were obtained from naturally
In Southeast Asia, especially in Laos, Cambodia, and Thai- infected cyprinoids fish captured from a freshwater reser-
land, infection with the liver fluke Opisthorchis viverrini is a voir in an endemic area of Khon Kaen, Northeast Thailand.
major health problem. In this region, the annual prevalence is Fresh fish were digested in 1% pepsin–HCl and incubated
more than six million (IARC 1994). In addition, O. viverrini at 37°C for 1 h, then filtered and precipitated with normal
saline in a sedimentation jar. Afterwards the metacercariae
were identified under a dissecting microscope.
T. Boonmars (*) : S. Boonjaraspinyo
Department of Parasitology, Faculty of Medicine,
Khon Kaen University, Animal infection
Khon Kaen 40002, Thailand
e-mail: [email protected] Six-week-old hamsters, gerbils, mice, and rats (five in each
group) were given 50 metacercariae by oral, intragastric
T. Boonmars
Liver Fluke and Cholangiocarcinoma Research Center, intubation. At 45 days post-infection, all of the animal
Faculty of Medicine, Khon Kaen University, groups were anesthetized with diethyl ether, weighed, and
Khon Kaen 40002, Thailand photographed. The liver, gall bladder, and extrahepatic bile
duct were carefully dissected and the worms collected. The
B. Kaewsamut
Northeast Animal Laboratory Center, Khon Kaen University, recovered worms were stained with carmine for morpholog-
Khon Kaen 40002, Thailand ical observation.
702 Parasitol Res (2009) 104:701–703

A D

B E

C F
Oral sucker

Ventral sucker

Vitelline
gland

Egg in uterus
Ovary
Testes

Recovered adult Recovered adult

worms of O.viverrini worms of O.viverrini
from hamsters from gerbils
Fig. 1 Gross anatomy of opisthorchiasis in hamster (a) and gerbil (d) livers. Recovered worms from hamsters (b and c) and gerbils (e and f)
stained with carmine. Magnifications ×4
Parasitol Res (2009) 104:701–703 703

The protocol was approved by the Animal Ethics Com- study O. viverrini infection in 1965 (Wyckoff et al. 1965).
mittee of the Faculty of Medicine, Khon Kaen University, Rabbit, dog, civet, and cat models were also used but not
Thailand (Ethical Clearance No AEKKU30/2551). widely (Healy 1970). Our results suggest that although the
hamster is popular, a gerbil model would be valid. The
reason for choosing a gerbil model would be: (1) gerbils are
Results more susceptible to O. viverrini infection than hamsters, (2)
the pathology of O. viverrini infection in gerbils is more
The examinations of the gross anatomy of the livers virulent than in hamsters, (3) the development of the
revealed that rat and mice livers were normal (data not parasite’s reproductive organs in gerbils is faster than in
shown), perhaps because the physio-anatomy of mice and hamsters, and (4) the EPG in feces in infected gerbils was
rats may not be optimal for the development of O. viverrini. greater than in hamsters.
For example, the rat does not have a gallbladder. There
were, however, changes in hamster livers, the gallbladder
was opaque and the hepatic bile ducts thickened (Fig. 1a). Conclusion
This result agrees with the previous reports (Wyckoff et al.
1965; Boonmars et al. 2007, 2008; Sripa 2003). Gerbil Gerbils may be the first choice for an animal model for
livers had the same changes to gross anatomy as observed research using eggs or excretory/secretory product or crude
in hamsters and the infection and pathological changes antigen because the parasite takes less time to develop in
more severe. The gerbil livers had a lot of nodules on the gerbils than hamsters. It is unclear why hamsters are the
surface of the liver and a more opaque gallbladder and more popular O. viverrini infection model.
thickened hepatic bile duct (Fig. 1d). The dilated gallblad-
der with green bile demonstrated obstruction of the hepatic Acknowledgements This research was supported by the Research
Strengthening Grant from National Center for Genetic Engineering
bile duct. Most of the gerbil gallbladders were larger than and Biotechnology (BIOTEC), National Science and Technology
the normal control. Development Agency (NSTDA). We thank the Department of
The number of recovered worms from gerbils was Parasitology, Liver Fluke and Cholangiocarcinoma Research Center,
significantly greater than from hamsters (P<0.05) (14±4.9 the Animal Experimental Unit, Faculty of Medicine, Khon Kaen
University, Department of Parasitology for their support and Mr.
and 30.25±7.22, respectively). The adult morphology shows Bryan Roderick Hamman for assistance with the English-language
the respective height×length of recovered adults from both presentation of the manuscript.
gerbils (Fig. 1e, f) and hamsters (Fig. 1b, c) was 5±0.12×
1.81±0.062 mm and 4.4±0.3×1.433±0.079 mm. The
difference was not statistically significant (P>0.05).
The sexual development of the worms was rapid, with References
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