M Magzoub

S E I Adam

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THE. JOtREAI OF PHARMACOlOGY AND EXPERIMENTAl. THERAPEUTiCS
Copyright 0 1978 by The American Society for Pharmacology and Experimental Therapeutics
Evaluation of Oral Therapy of Mansonial
Schistosomiasis in the Sudan Using Single and
Multiple Doses of Hycanthone and Oxamniquine
Drug&
M. MAGZOUB and S. E. I. ADAM
Departments of Parasitology and Clinical Studies, Faculty of Veterinary Science, University of Khartoum, Sudan
Accepted for publication February 24, 1978
ABSTRACT
Magzoub, M. and S. E. I. Adam: Evaluation of
oral therapy of mansonial schistosomiasis in the
Sudan using single and multiple doses of hycan-
thone and oxamniquine drugs. J. Pharmacol.
Exp. Ther. 206: 382-387, 1978.
Single doses of 20 mg/kg of hycanthone or
oxamniquine or a combination of 20 mg/kg of
each drug, given to Nile rats ( Arvicanthus nib-
ticus) infected with Schistosoma mansoni, did
not produce significant changes in the parasitic
fauna or in the hepatic damage induced by the
parasite. Three daily doses of 20 mg/kg of
these drugs alone or in combination reduced
considerably the intensity of parasitic infection
and caused a marked recovery from the tissue
damage induced by parasitic invasion. The pro-
portions of immature eggs, as compared with
the mature ones, were 1 to 1 1 .5 in presence of
hycanthone and 1 to 8 in presence of oxamni-
quine. There were no signs of immature eggs
when the two drugs were used in combination.
All living worms and nearly almost all immature
eggs disappeared from the tissues of the treated
animals examined 1 0 and 20 days after comple-
tion of treatment with hycanthone and oxamni-
quine alone as well as in combination. Gross
and microscopic examination of the animals
given both drugs in combination revealed the
absence of worms and eggs in the portal tracts
and reduced hepatocellular necrosis and schis-
tosome pigment in the Kupfter cells. In the
treated rats there was less of a decrease in the
glycogen content of the hepatocytes but aggre-
gation of mononuclear cells was found in the
portal areas. Splenomegaly associated with hy-
perplasia and granulomatous splenitis was seen
in the untreated infected rats, but not in the
infected rats given a combination of hycanthone
and oxamniquine.
The present experiments were concerned with
the treatment of intestinal schistosomiasis in the
Nile rat (Arvicanthus niloticus) with single,
multiple and mixed doses of hycanthone and
oxamniquine. The criteria used for the evalua-
tion of these antischistosomal agents included
the susceptibilities to drugs displayed by adult
schistosomes and the effect of treatment on
Received for publication January 5, 1977.
I This work was supported by a grant from the National
Council for Research, Khartoum.
pathological changes in different organs caused
by bilharzia infection.
Materials and Methods
One hundred Nile rats (A. niloticus) of both sexes
and weighing 40 to 60 g each were used for the exper-
imental study. These were randomly grouped. a) Four
unirifected rats were killed by a blow on the head
during the 9th week after the start of the experiment.
b) Ten rats, each infected percutaneously with 200
cercariae of our local (Sudan) strain of Schistosoma
7nansoni, were killed in subgroups of at least 3 at 49
1978 Oral Therapy for Schistosomiasis 383
days (bi), 56 days (b2) and 63 days (bJ) from the date
of infection. c) Twenty-eight infected rats were dosed
orally, 8 weeks after infection, with hycanthone given
in a dose of 20 mg/kg. Hycanthone was dissolved in
distilled water and a single dose was given to 3 rats
(ci); three daily doses to 25 rats (c2), killed after 3
(c2a), 10 (c2b) and 20 (c2c) days after the end of
treatment. d) Twenty-eight infected rats were treated
orally, 8 weeks after infection, with oxamniquine (20
mg/kg) which was prepared in the same way as hy-
canthone; a single dose was given to 3 rats (dl) and
three daily doses were given to 25 rats (d2) killed after
3 (d2a), 10 (d2b) and 20 (d2c) days after the completion
of treatment. e) Thirty infected rats were treated, 8
weeks after infection, with a combination of hycan-
thone and oxamniquine, each in a dose of 20 mg/kg; a
single dose of the combined drugs was given to 4 rats
(el), one of which died during treatment. Twenty-five
other rats were given three daily doses ofthe combined
drugs (e2) and killed after 3 (e2a), 10 (e2b) and 20
(e2c) days from the end of treatment.
Histological and histochemical methods. Spec-
imens of liver, intestines, spleen, kidneys, lungs and
heart were fixed in 10% formol-saline, embedded in
paraffin wax and sectioned at 5 m. All sections were
stained with hematoxylin and eosin (H&E) and se-
lected sections were stained by Prussian blue, periodic
acid-Scuff (PAS) with and without prior incubation
with diastase and Schmorl’s method for lipofuscin.
Results
Parasitulogical findings. Ten rats were sac-
ri.ficed, three 49 days, four 56 days and three 63
days after exposure to 200 cercariae of S. man-
Soni. In all rats examined, the number of the
immature eggs in the tissues of the intestine was
greater than the mature ones. Forty-nine days
after infection, the immature eggs represented
76% and the mature eggs 15% of the total; 56
days after infection, the immature eggs repre-
sented 77% and the mature eggs 15% of the total;
63 days after infection, the percentages of im-
mature and mature eggs were 71 and 17%, re-
spectively. There were 9% dead eggs 49 days
after infection, 8% 56 days after infection and
22% 63 days after infection.
The worms from the liver and mesenteric
vessels of the infected and treated animals were
recovered by the perfusion technique of Smith-
em and Terry (1965) and were counted during
postmortem examination of each rat. The data
are presented in table 1. The average number
and sex of the worms found in the body of each
infected rat in the first group was 20 males and
16 females in the mesenteric vessels and 14 males
and 10 females in the liver; in the second group
18 females and 22 males were recovered from
the mesenteric vessels and 16 males and 14
females from the liver; in the third group, killed
63 days after infection, 22 females and 26 males
were recovered from the mesenteric vessels and
12 females and 18 males from the liver.
Hycanthone experiments. Three days after
the completion of treatment, 8 rats were killed
and examined for the presence of adult worms
and eggs of S. mansoni in the tissues of the
intestine; the average number of worms found in
the liver of each rat was 2 males and 1 female
and in the mesenteric vessels 4 males and 3
females; of the eggs found, 2% were immature,
23% were mature and 75% were dead.
Ten days after completion of treatment, 7 rats
were examined and the results revealed a corn-
plete absence of living worms in the liver and
intestines of the treated animals. Examination
of the eggs showed 1% were immature, 53% were
mature and 46% were dead.
Ten infected rats were examined for living
worms and eggs of the schistosome parasite 20
days after treatment. No worms were found in
the liver and intestines. Examination of the eggs
in the tissues of the intestine revealed 53% were
mature and 47% were dead; there was a complete
absence of immature eggs.
Oxamniquine experiments. Three days
after the completion of the treatment with ox-
amniquine, 8 rats were examined for living
worms and eggs of the schistosome parasite.
Fourteen males and 10 female worms were found
in the liver and intestines of these animals. Ten
days after completion of treatment, 5 males and
3 females were detected in the tissues ofthe liver
and intestines of 8 rats. Twenty days after the
treatment, 9 rats were examined; living worms
in the liver and intestines were absent.
Examination of eggs in the intestines of these
animals 3 days after the end oftreatment showed
3% were immature, 23% were mature and 74%
were dead. Ten and 20 days after the end of
treatment, the percentages were 1 and 0% im-
mature, 34 and 31% mature and 65 and 69%
dead, respectively.
Combined administration of hycanthone
and oxamniquine. Three days after the end of
treatment with the two drugs in combination, 5
rats were examined for living worms and eggs of
the schistosome parasite. Four males and 2 fe-
males were picked from the liver and intestines
of these rats. Ten and 20 days after the treat-
ment, no living worms were found in the liver
and intestines of the 20 animals examined.
The oogram study showed 0% immature, 29%
mature and 71% dead eggs 3 days after the end
of treatment; 10 and 20 days after completion of
384 Magzoub and Adam VoL 206
treatment the immature eggs were absent, the Infected rats. There were irregular nodules
mature were 18 and 1% and the dead eggs 82 and and fatty change. Many enlarged portal tracts
99%, respectively. showed egg granulomata (fig. 1). The cellular
Pathological changes. Liver. The effects of infiltrations consisted of round cells, eosinophil
hycanthone and oxamniquine treatments on the leukocytes, mononuclear cells and fibroblasts.
liver changes produced by S. mansoni infection There was fatty cytoplasmic vacuolation, focal
in the Nile rat are summarized in table 2. necrosis and depletion of glycogen in the par-
TABLE 1
Efficacy of Hycanthone and Oxamniquine In the treatment of S. mansoni infection in the Nile rat
Average No. of Worms in
No. of
No. of Rats Age of In- Dose of Daily Mesenteric
Group . . Liver
per Group fection Drug Doses of veins
Drug
9  9
days mgi kg
Uninfected control
A 4
Infected control
Bi 3 49 60 48 42 30
B2 4 56 88 72 64 56
B3 3 63 78 66 54 36
Hycanthone
Cl 3 56 20 1 46 28 26 18 3 days post-
treatment
C2a 8 56 20 3 32 24 16 8 3days post-
treatment
C2b 7 56 20 3 1 0 days post-
treatment
C2c 10 56 20 3 2odays post-
treatment
Oxamniquine
dl 3 56 20 1 42 24 24 16
d2a 8 56 20 3 8 6 6 4 3 days post-
treatment
d2b 8 56 20 3 3 2 2 1 10 days post-
treatment
d2c 9 56 20 3 20 days post-
treatment
Hycanthone + oxamni-
quine
el 4 56 20 1 20 12 8 6
e2a 5 56 20 3 4 1 2 1 3 days post-
treatment
e2b 10 56 20 3 lOdays post-
treatment
e2c 10 56 20 3 2odays post-
treatment
TABLE 2
Assessment of hepatic damage in infected Nile rats treated with hycanthone and oxamniquine
No of Uver Changes#{149}
Doseof
Drug Daily
Drug Fatty . Loss of gly- Schistoso-
Doses Granulomas Necrosis
change cogen ml pigment
(mgi kg)
Infected control + + + + + + + + + +
Hycanthone 20 1 + + + + + + +
Hycanthone 20 3 + + (-) + (-)
Oxamniquine 20 1 + + + + + + +
Oxamniquine 20 3 + + (-) + (-)
Hycanthone + oxamniquine 20 3 + + (-) + (-)
. +-#{248} + ± + . increasing severity of lesions: (-), absence of lesions.
FIg. 1. Rat liver 9 weeks after infection with 200 cercariae of S. mansoni: egg granuloma. (H&E, x235.)
1978 Oral Therapy for Schistosomiasis 385
enchyma. Kupffer cells in the periphery of the
lobule and the subcuticle ofthe worms contained
schistosomal pigment which did not stain with
either Schmorl’s or Prussian blue methods.
Hycanthone-treated rats. The numbers of
granulomas in the portal vessels and the levels
of glycogen and fatty change were slightly re-
duced in the liver of rats which received single
doses of hycanthone. After three doses, there
was no evidence of schistosomal pigment in
Kupffer cells. The number of granulomas in the
portal areas was considerably reduced and many
portal tracts showed neither worms nor ova.
There was infiltration of mononuclear cells and
eosinophil leucocytes in the portal tracts (fig. 2).
Focal necrosis disappeared and the glycogen
content was increased in the parenchyma.
Oxamniquine-treated rats. The livers of rats
in the single dose group did not differ signifi-
cantly from the infected controls. After three
doses, there was neither hepatic necrosis nor
deposition of schistosomal pigment in Kupffer
cells. The number of granulomas was reduced
and the hepatocytes showed an increase in gly-
cogen content.
Hycanthone and oxamniquine treatments.
The lesions in the liver resembled those found
previously in rats given three doses of either
hycanthone or oxamniquine. The numbers of
granulomas in the portal areas and of the worms
and ova were very low compared to the infected
controls.
Other tissues. The findings in other tissues of
infected rats were infiltration of the intestinal
lamina propria with inflammatory cells, pulmo-
nary congestion and/or edema and splenomeg-
ely (fig. 3). Splenomegaly was also seen in rats
which received single doses of oxamniquine.
After three doses of hycanthone or oxamniquine,
there were few such changes.
Discussion
To fmd more suitable anthelminthics for use
against Schistosoma mansoni, attention was
turned to orally administered drugs. Magzoub
and Maegraith (1969) showed that niridazole
was very active in reducing the motility of the
Egyptian strain of S. mansoni through the in-
hibition of the exogenous glucose uptake from
blood and the subsequent depletion of glycogen
in the tissues of the treated worms. Magzoub
and Adam (1973) tested lucanthone hydrochlo-
ride for its activity against the Sudan strain of
S. mansoni in albino mice and found that the
drug was most active in killing the worms and
preventing the development of hepatic damage
when given in 10 daily doses of 100 mg/kg.
In this paper, we tested hycanthone and ox-
amniquine against S. mansoni infection in Nile
. 
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386 Magzoub and Adam Vol. 206
FIg. 2. Rat liver after treatment with three daily doses of hycanthone: absence of egg granuloma and infiltration
of mononuclear cells in the portal tracts. (H&E, x235.)
‘Lr
Fig. 3. Splenomegaly in a Nile rat 9 weeks after
infection with 200 cercariae of S. mansoni.
rats (Arvicanthus niloticus). After single doses
of either hycanthone or oxamniquine, there was
no significant reduction in the number of adult
worms in the liver and the intestines, no change
in the oogram of the intestines and no significant
decrease either in the number of granulomas or
of hepatocellular necrosis. These findings mdi-
cate that the administration of both drugs at
single oral doses did not produce any significant
effect on the worm load and on the hepatic
damage induced by the parasite.
The oral administration of hycanthone, ox-
amniquine and their combination in three daily
doses of 20 mg/kg proved to be more effective.
In the presence of hycanthone, infected rats
(killed after 10 days from treatment) showed no
living worms in the tissues of the liver and
intestines and a complete absence of immature
eggs in the intestines. Ten days after the end of
treatment with oxamniquine, no living worms
were seen in the tissues of the treated animals
and the oogram of the intestines showed the
presence of 1% immature eggs as compared with
77% in the absence of treatment. Animals
treated with mixed doses of both drugs and
examined 10 days after treatment showed no
living parasites and no immature ova in the
tissues of the intestines.
However, our pathological investigations
showed that the administration of hycanthone,
oxamniquine or their combination in three daily
doses proved to be an effective treatment for the
removal of S. mansoni as indicated by the ab-
sence of the parasites and ova from the portal
tracts, of hepatocellular necrosis and schisto-
some pigment from Kupffer cells. The glycogen
content was increased in the hepatocytes and
aggregates of mononuclear cells were found in
the portal areas. Schiller and Haese (1973) re-
References
1978 Oral Therapy for Schistosomiasis 387
HAE.SE, W. H. AND BUEDING, E.: Long.term hepatocellular
effects of hycanthone and of two other anti.schistosomal
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weeks after elimination of the worms by treat-
ment. The carcinogenicity of hycanthone in
Swiss-Webster strain of mice has been demon-
strated by Haese and Beuding (1976).
Hepatosplenomegaly is a constant finding in
S. mansoni infection in man and is accompanied
by marked eosinophilia (Schiff, 1963). We did
not observe splenomegaly in albino mice in-
fected with a Sudan strain of S. mansoni (Mag-
zoub and Adam, 1973, 1974a,b). The results re-
corded in this paper demonstrated the presence
of splenomegaly in the Nile rat infected with a
Sudan strain of S. mansoni and this was associ-
ated with hyperplasia and granulomatous splen-
itis.
Acknowledgments
We thank Mr. H. Ibrahixn and Mr. A. H. Saad for technical
assistance and Mr. E. M. Abu Ganaya for photography.
drugs in mice infected with Schistosoma mansoni. J. Phar
macol. Exp. Ther. 197: 703-713, 1976.
MAGZOUB, M. AND ADAM, S. E. I.: Effect of lucanthone
hydrochloride on the hepatic lesions induced by Sudan
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MAGZOUB, M. AND ADAM, S. E. I.: Experimental infection and
hepatic changes in mice infected with Schistosoma mansoni
(Sudan strain). J. Pathol. 113: 47-52, 1974a.
MAGZOUB, M. AND ADAM, S. E. I.: A comparative study of the
liver damage and worm burden following infection with
Schistosoma mansoni from northern and southern Sudan.
Br. J. Exp. Pathol. 55: 260-276, 1974b.
MAGZOUB, M. AND MAEGRAITH, B. G.: Glycolytic activities of
Schistosoma mansoni and the mode of action of antischis.
tosomal agents (Ambilhar and Astiban). Ann. Trop. Med.
Parasitol. 63: 377-391, 1969.
SCHIFF, L.: Diseases of the Liver, 2nd ed., Pitman Medical
Publishing Comp. Ltd., Philadelphia, 1963.
SCHILLER, E. S. AND HAE.SE, W. H.: Histologic processes of
dealing in hepatic injury due to ova ofSchistosoma mansoni
in mice following curative chemotherapy. Amer. J. Trop.
Med. Hyg. 22: 211-214, 1973.
SMITHERS, S. R. ANDTERRY, R. J.: The infection of laboratory
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covery of the adult worms. Parasitology 55: 695-700, 1965.
Send reprint requests to: Dr. S. E. I. Adam, Department of
Veterinary Clinical Studies, University ofKhartoum, P.O. Box
32, Khartoum, North Sudan.


Evaluation of Oral Therapy of Mansonial Schistosomiasis in the Sudan Using Single and Multiple Doses of Hycanthone and Oxamniquine Drug&amp;

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Evaluation of Oral Therapy of Mansonial Schistosomiasis in the Sudan Using Single and Multiple Doses of Hycanthone and Oxamniquine Drug&

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