Article available at http://www.parasite-journal.org or http://dx.doi.org/10.1051/parasite/200108s2195
EFFICACY OF FLUBENDAZOLE AND ALBENDAZOLE
AGAINST TRICHINELLA SPIRALIS IN MICE
CHUNG M.S.*, J O O K.H.*, QUAN F.S.*, K W O N H.S.* & C H O S.W.*
in mice. Lopez-Garcia et al. ( 1 9 9 7 ) compared
Summary:
T. spiralis
Efficacy of flubendazole and albendazole against Trichinella
spiralis in mice were studied. ICR mice were experimentally
infected with Trichinella spiralis and treated with either
flubendazole (FBZ) or albendazole (ABZ) at four different stages of
the parasite life-cycle. Oral administration of either FBZ or ABZ at
2 0 mg/kg and 5 0 mg/kg on 2 h, 8 h and 2 4 h (pre-adult
stage) after infection eliminated 9 4 . 7 - 100 % of adults as
determined at necropsy on day 7 post infection (p.i.) and 9 6 . 9 ~
100 % of larvae on day 4 5 p.i. FBZ was more effective than
ABZ against adult T. spiralis (at 2 to 6 days p.i.), when treated
with a dosage of 2 0 mg/kg for 5 consecutive days (99.4 % and
4 6 . 0 % reduction with respect to the control group). Against
migrating larval T. spiralis, FBZ was more effective than ABZ at
2 0 mg/kg for five consecutive days (on days 11̰15 p.i.), and
the reduction rate of recovered larvae were 9 9 . 6 % (FBZ) and
80.8 % (ABZ) respectively. FBZ was more effective against early
encapsulated larval T. spiralis (at 21 to 25 days p.i.), than ABZ
when both were given at 2 0 mg/kg for five consecutive days
(99.8 % and 4 5 . 4 % reduction, respectively). In conclusion,
flubendazole was more effective than albendazole against adult
and parenteral stages of Trichinella spiralis in mice.
the effect o f a l b e n d a z o l e and ricobendazole (alben-
KEY WORDS : Trichinella spiralis, flubendazole, albendazole, efficacy, mice.
T
d a z o l e s u l p h o x i d e ) against enteral a n d parenteral
stages o f T. spiralis
in m i c e . H o w e v e r , there has b e e n
f e w reports that effectiveness o f flubendazole against
different stages o f Trichinella
spiralis
in mice.
T h e objective o f this study w a s to evaluate the effect
o f t w o benzimidazole c o m p o u n d s , flubendazole ( F B Z )
and a l b e n d a z o l e ( A B Z ) o n different stages in the life
cycle o f Trichinella
spiralis
infection in mice.
MATERIALS AND METHODS
PARASITE
Y
amagata strain o f Trichinella
spiralis
w a s used
for the experiments. This strain has b e e n maintained in our laboratory by passage through rat
since 1993.
EXPERIMENTAL ANIMALS
richinellosis is o n e o f the most widespread helm i n t h i c z o o n o s e s . In K o r e a , t h r e e c a s e s
h u m a n infection b y Trichinella
of
were
spiralis
first confirmed by detecting encysted larvae in the biopsied muscle in D e c e m b e r
1997. T h e patients
were
treated with flubendazole a n d a l b e n d a z o l e for 1 5 - 3 0
days ( S o h n et al., 2 0 0 0 ) .
S o m e reports have demonstrated the chemotherapeutic
effectiveness o f benzimidazole c o m p o u n d s in the treatment o f Trichinella
spiralis
infection in m i c e (Camp-
bell & Cuckler, 1 9 6 4 ; Duckett & D e n h a m , 1 9 7 0 ; Spaldonova
et al.,
1974; Fernando & Denham,
Lopez-Garcia et al.,
1976;
1 9 9 7 ) . It w a s demonstrated that
m e b e n d a z o l e w a s highly effective against the immature enteral p h a s e o f trichinosis in m i c e ( F e r n a n d o &
D e n h a m , 1 9 7 6 ; McCracken, 1 9 7 8 ) . McCracken ( 1 9 7 8 )
F e m a l e ICR m i c e weighing 2 5 - 3 0 g w e r e used. Each
group w a s consisted o f six animals w h i c h w a s orally
infected with 2 5 0 larval T.
spiralis.
D R U G S AND TREATMENTS
Infected m i c e w e r e treated with either flubendazole
( F B Z ) o r a l b e n d a z o l e ( A B Z ) at four different stages o f
the parasite life-cycle: pre-adult, adult, migrating larvae
and early encapsulated muscle larvae. Oral administration o f either FBZ or ABZ at 20 mg/kg and 50 mg/kg
on 2 h, 8 h and 24 h (pre-adult stage), 72 h (adult
stage), and 2 0 mg/kg for five consecutive days (adult,
migrating and early encapsulated larval stages).
R E C O V E R Y O F WORMS
reported that a l b e n d a z o l e w a s m u c h m o r e effective
T h e n u m b e r o f enteral Trichinella
against pre-adult a n d had a partial effect o n adult
mice w a s estimated seven days after exposure to infec-
in e a c h group o f
tion. At necropsy, e a c h m o u s e w a s e x a m i n e d indivi* Department of Parasitology, Korea University College of Medicine,
136-705, Seoul, Korea.
Tel.: 822-920-6178 - Fax: 822-924-4905
e-mail: [email protected]
Parasite, 2001, 8, S195-S198
dually for adult w o r m s from gut. All the mice treated
with drugs against migrating a n d early encapsulated
muscle larval stage w e r e killed 45 days post infection
(p.i.) and the number o f diaphragm larvae was counted.
X ICT, August 2 0 0 0
th
S195
CHUNG M.-S., JOO K.-H., QUAN F.-S., KWON H.-S. & CHO s.-w.
same dosages w e r e only partially active at 24, 4 8 h p.i.
RESULTS
( T a b l e s I, II).
E F F E C T O F T H E D R U G S O N PRE-ADULT AND ADULT
STAGE
C
T. SPIRALIS
F B Z s h o w e d similar anthelmintic acitivity against the
w o r m s in intestinal p h a s e . W h e n the infection w a s 8 h
old, a single d o s e o f 20 and 50 mg/kg r e m o v e d all o f
onsistent with the earlier report o f McCracken
the Trichinella
( 1 9 7 8 ) , albendazole was highly active against the
later ( T a b l e III, IV). At 48, 72 h after inoculation, acti-
as determined at n e c r o p s y s e v e n day
w o r m s in the immature enteral p h a s e o f trichi-
vity w a s not c o m p l e t e l y curative, at b o t h
dosages
nellosis. A single oral d o s e o f 20 and 50 mg/kg 2 h
( T a b l e III, IV). F B Z and ABZ w e r e m o r e
effective
or 8 h after inoculation totally eradicated the infection
against pre-adult Trichinella
as determined b y examination o f the gut for adult
infected
w o r m s on day 7 p.i. ( T a b l e s I, II). H o w e v e r , as worms
2 0 mg/kg for five c o n s e c u t i v e days, there was signifi-
matured, their susceptibility to ABZ diminished; the
cant difference in the reduction in w o r m b u r d e n bet-
mice were
Adult*
control mice
treated mice
2
8
24
48
72
h
h
h
h
H
than adult w o r m s . W h e n
treated with
the
dosage
of
Larvae**
Mean No.
worms/mouse
Efficacy
of treatment
Mean No.
worms/mouse
1 2 6 . 5 ± 21.5
0
0
0
93 ± 9.2
59.7 ± 6.1
_
100 %
100 %
100 %
26.5 %
52. 8 %
1,022 ± 17
0
0
0
128 ± 5.5
53 ± 4.5
Efficacy
of treatment
_
100
100
100
87.5
94.8
%
%
%
%
%
* Collection at 7 dpi in intestine.
** Collection at 45 dpi in 0.1 g of diaphragm.
Table I. - Number of T. spiralis worms recovered from the intestine and diaphragm of mice treated with 20 mg/kg single dose of
albendazole at various time after infection.
Adult*
Mean No.
worms/mouse
control mice
treated mice
2h
8 h
24 h
48 h
72 H
126.5 ± 21.5
0
0
0.67 ± 0.47
20 ± 7.8
41 ± 21
Larvae**
Efficacy
of treatment
Mean No.
worms/mouse
_
1,022 ± 17
0
0
0
29.3 ± 13.9
12.7 ± 11.5
100
100
99.5
84.2
67.6
%
%
%
%
%
Efficacy
of treatment
_
100
100
100
97.2
98.7
%
%
%
%
%
* Collection at 7 dpi in intestine.
** Collection at 45 dpi in 0.1 g of diaphragm.
Table II. - Number of T. spiralis worms recovered from the intestine and diaphragm of mice treated with 50 mg/kg single dose of
albendazole at various time after infection.
Adult*
Mean No.
worms/mouse
control mice
treated mice
1
8
24
48
72
h
h
h
h
H
126.5 ± 21.5
2 ± 1.6
0
6.67 ± 5.2
46 ± 12.7
78.5 ± 8.5
Larvae**
Efficacy
of treatment
98.4
100
94.7
63.6
37.9
%
%
%
%
%
Mean No.
worms/mouse
1,022 ± 17
0
0
32.3 ± 3.4
56.5 ± 36.5
34.3 ± 4.0
Efficacy
of treatment
100
100
96.9
94.5
96.7
%
%
%
%
%
* Collection at 7 dpi in intestine.
** Collection at 45 dpi in 0.1 g of diaphragm.
Table III. - Number of T. spiralis worms recovered from the intestine and diaphragm of mice treated with 20 mg/kg single dose of flubendazole at various time after infection.
S196
X ICT, August 2000
th
Parasite, 2 0 0 1 , 8, S195-S198
PATHOLOGY AND TREATMENT
Adult*
control mice
treated mice
2
8
24
48
72
h
h
h
h
H
Larvae**
Mean No.
worms/mouse
Efficacy
of treatment
Mean No.
worms/mouse
126.5 ± 21.5
0
0
0.33 ± 0.47
1 ± 1.41
16 ± 4
100%
100 %
99.7 %
99.2 %
87.3 %
1.022 ± 17
0
0
0
23 ± 8.4
5.3 ± 1.2
Efficacy
of treatment
_
100
100
100
97.7
99.5
%
%
%
%
%
* Collection at 7 dpi in intestine.
** Collection at 45 dpi in 0.1 g of diaphragm.
Table IV. - Number of T. spiralis worms recovered from the intestine and diaphragm of mice treated with 50 mg/kg single dose of flubendazole at various time after infection.
Adult*
Individual counts
Mean
Larvae**
Control
ABZ
FBZ
Control
ABZ
98
126
136
118
106
130
91
43
70
72
44
1
0
1
1
1
0
2,131
2,308
1,221
1,267
1,547
98
218
61
131
126
118.5
64.3
0.67
1,694.8
126.8
0.5
46.0 %
99.4 %
92.5 %
99.97 %
Efficacy of treatment
64
FBZ
0
0
0
1
0
* Collection at 7 dpi in intestine.
** Collection at 45 dpi in 0.1 g of diaphragm.
Table V. - Number of T. spiralis worms recovered from the intestine of mice treated with albendazole and flubendazole at a dosage of
20 mg/kg/day for 5 consecutive days ( 2 - 6 days pi; adult stage).
w e e n F B Z ( 9 9 . 4 % ) and ABZ ( 4 6 . 0 % ) ( T a b l e V ) .
counted. FBZ was more effective than A B Z ; 99-8 % and
T h e s e results p r o m p t e d a further examination o f the
4 5 . 4 % reduction, respectively ( T a b l e VII).
c h a n g e s in drug susceptibility during the
parasite's
Results also s h o w that w h e n given as a series o f oral
doses, F B Z is m o r e efficacious against the invasive
period o f maturation in the gut.
p h a s e than ABZ.
E F F E C T O F THE DRUGS O N MIGRATING AND EARLY
ENCAPSULATED LARVAL STAGE
T. SPIRALIS
Larvae
Despite the decline in drug sensitivity, two drugs w e r e
further tested for effectiveness against the invasive phase
Control
ABZ
FBZ
1.622
1,278
1,521
2,458
2,308
1,778
384
405
449
218
301
5
30
3
6
1827.5
351.4
7.3
80.8 %
99.6 %
o f trichinellosis because o f its special clinical importance.
Results are tabulated in Table VI. Mice treated for five
Individual counts
days with FBZ yielded a m e a n o f 7.3 larvae, which is
99.6 % less than the control m e a n o f 1,827.5. T h e average number o f larvae in the mice treated with ABZ was
351.4, representing a reduction o f 8 0 . 8 % as c o m p a r e d
to the untreated controls (Table VI).
Mean
T o determine the efficacy o f drugs against early e n c a p sulated m u s c l e larvae, six m i c e r e c e i v e d FBZ
and
other group o f six m i c e ABZ at 20 mg/kg on e a c h day
from the 21st day p.i. during five successive days. All
the m i c e w e r e killed 20 days after the last day o f treatment and the n u m b e r o f larvae from diaphragm w e r e
Parasite, 2001, 8, S195-S198
Efficacy of treatment
Table VI. - Number of T. spiralis larva recovered at 45 dpi from the
diaphragm of mice treated with albendazole and flubendazole at a
dosage of 20 mg/kg/day for five consecutive days (11 - 15 dpi;
migrating larval stage).
X ICT, August 2000
th
II
0
S197
C H U N G M.-S., J O O K . - H . , Q U A N F . - S . , K W O N H . - S . & C H O S . - W .
larvae recovered from the host musculature. In this
Larvae
Control
Individual counts
paper, administration o f flubendazole at migrating larval
FBZ
ABZ
1,622
1,278
1,521
513
839
1,662
0
1s
2,458
998
0
Lopez-Garcia et al., 1997). Administration o f m e b e n d a -
2,308
979
3
0
zole, chemically similar to flubendazole, against encysted
0
1,778
Mean
1827.5
Efficacy of treatment
998.2
3.5
45.4 %
99.8 %
Table VII. - Number of T. spiralis larva recovered at 45 dpi from
the diaphragm of mice treated with albendazole and flubendazole
at a dosage of 20 mg/kg/day for five consecutive days (21 ~25 dpi;
early encapsulated muscle larval stage).
m e b e n d a z o l e a n d albendazole w e r e highly effective
against encysted larvae (Fernando & Denham, 1 9 7 6 ;
larval Trichinella
the present study, oral administration o f F B Z eliminated
9 9 . 8 % o f larvae o n day 4 5 p.i. In conclusion,
fluben-
dazole w a s m o r e effective than albendazole
against
adult a n d parenteral stages o f T. spiralis
the enteral and parenteral phases of trichinosis in mice.
h e Triehinella/mouse
model has b e e n widely
u s e d to assay anthelmintic effectiveness o f b e n zimidazole derivatives (Campbell & Blair, 1974 ;
& Denham,
McCraken & Taylor,
Lopez-Garcia et al,
1976 ; McCracken, 1 9 7 8 ;
1 9 8 0 ; McCracken
et al, 1 9 8 4 ;
1 9 9 7 ) . T h e present findings, toge-
ther with the reports cited earlier, indicate that m e m b e r s o f this c h e m i c a l class are m u c h m o r e effective
against the pre-adult than adult w o r m .
In t h e present study, F B Z was m u c h m o r e active in eliminating adult
in mice.
REFERENCES
J
Fernando
reduced the larvae recovered from the
host muscle b y 99.7 % (Fernando & D e n h a m 1976). In
CAMPBELL W.C. & CUCKLER A.C. Effect of Thiabendazole upon
DISCUSSION
T
reduced the number o f larvae obtained from
T. spiralis
diaphragm in mice b y 9 9 . 6 %. It w a s reported that
T. spiralis.
ABZ, o n the other hand,
Parasitol,
1 9 6 4 , 50. 4 8 1 - 4 8 6 .
CAMPBELL W.C. & BLAIR L.S. Chemotherapy of Tricbinella
ralis infections (a review). Exp. Parasitol.,
spi-
1 9 7 4 , 35. 3 0 4 - 3 3 4 .
DUCKETT M.G. & DENHAM D.A. The effect of Cambendazole
on Trichinella
1970,
spiralis infections in mice. J. Helminthol.,
44, 2 1 1 - 2 1 8 .
FERNANDO S.S.E. & DENHAM D.A. The effects of mebendazole
fenbendazole on Trichinella
1976,
spiralis in mice. J. Parasitol.,
62, 8 7 4 - 8 7 6 .
LOPEZ-GARCIA M.L., TORRADO-DURAN S., TORRADO-DURAN J . ,
MARTINEZ-FERNANDEZ A.R. & BOLAS-FERNANDEZ F. Albenda-
these differences m a y b e d e p e n d e n t as Campbell &
zole versus ricobendazole (albendazole-sulphoxide)
against enteral and parenteral stages of Trichinella
spiralis
in mice. Int. J. Parasitol.. 1 9 9 7 , 22, 7 8 1 - 7 8 5 .
C u c k l e r ( 1 9 6 4 ) with t h i a b e n d a z o l e a n d D u c k e t t &
GABRYEL P., GUSTOWSKA L. & BLOTNA M. Morphology and his-
D e n h a m ( 1 9 7 0 ) with c a m b e n d a z o l e found that high
c o m p l e t e elimination o f adult worms. A s e x difference
tochemistry of muscle in Trichinella-infected
rat treated
with thiabendazole. Proceedings Third International Conference on Trichinellosis, Nov. 2 - 4 , 1 9 7 2 .
was reported for piperazine, which was said to b e more
GERWTL C , PAWLOWSKI Z., KOCIECKA W. & CHODERA L. Probable
s h o w e d a partial effect o n adult worms. However, all
level or long term administration o f c o m p o u n d caused
potent against male
Trichinella
than against
female
(Campbell & Blair, 1 9 7 4 ) . Although adult w o r m s w e r e
less removed, larvae w e r e highly reduced. F B Z and
ABZ were more effective against female than male (data
not s h o w n ) . Thiabendazole was reported to cause c o m plete cessation o f larval production b y female T. spiralis
in mice (Campbell & Cuckler 1 9 6 4 ) . T h e related
c o m p o u n d c a m b e n d a z o l e w a s also reported to suppress larval production (Duckett & D e n h a m , 1 9 7 0 ) .
Benzimidazoles acted o n parenteral forms
Tricbinella
(Campbell & Blair, 1 9 7 4 ) . Despite a low degree o f
absorption from the gut, mebendazole was highly active
against parenteral as well as enteral forms o f
Trichinella
in rats (Campbell & Blair, 1974) a n d mice (McCracken,
1978). McCracken (1978) demonstrated that gavage administration o f either mebendazole or albendazole during
the invasive phase o f Tricbinella
infection significantly
reduced b y 9 6 % and 6 7 %, respectively, the number o f
S198
sterilization of Trichinella spiralis by thiabendazole: Further clinical observations of human infections. Proceedings
Third international Conference on Trichinellosis, Nov. 2 4, 1 9 7 2 .
MCCRACKEN R.O. Efficacy of mebendazole and albendazole
against Tricbinella spiralis in mice. J Parasitol., 1 9 7 8 ,
64.
214-219.
MCCRACKEN R.O. & TAYLOR D . D . Mebendazole therapy of
parenteral trichinosis. Science,
zole : regime-dependence expression of drug efficacy against
Trichinella spiralis. Int. J. Parasitol., 1 9 8 4 , 14, 2 7 7 - 2 8 1 .
SPALDONOVA R., TOMASOVICOVA O. & CORBA J . The influence
of mebendazole on the course of Trichinella spiralis infection in mice. Proceedings of the Third International
Congress of Parasitology, 1 9 7 4 , Munich, 6 7 5 - 6 7 6 .
SOHN W.M., KIM H.M., CHUNG D . I . & YEE S.T. The first human
case of Tricbinella
spiralis infection in Korea. Kor.J.
Para-
sitol., 2 0 0 0 , 38, 1 1 1 - 1 1 5 .
X ICT, August 2 0 0 0
th
1 9 8 0 , 207, 1 2 2 0 - 1 2 2 2 .
MCCRACKEN R.O., NIERSTE D.M., Moss J . & GARCIA A. Oxfenda-
P a r a s i t e , 2001, 8, S195-S198