Abdul-Hussein H Awad, Alim A-H Yacoub, Sabeeh H Al-Mayah. Dynamic Aspects of Schistosoma Haematobium Infection as Experimental Model. Medical Journal of Basra University 1995;13(1&2):21-30

1. MJBU, Vol 13. No. 1&2, 1995
21
DYNAMICAL ASPECTS of SCHISTOSOMA HAEMATOBIUM
INFECTION: AS EXPERIMENTAL MODEL.
Abdul- Hussein H Awad, Alim A- H Yacoub, Sabeeh H AL-Mayah
Abdul- Hussein H Awad, PhD, Assistant Professor, College of Education, University of Basrah,
Iraq.
Alim A H Yacoub, PhD, MECM, Assistant Professor, College of Medicine, University of
Basrah, Basrah, Iraq.
Sabeeh H AL-MaYah, MSc, Assistant lecturer, College of Education, University of Basrah,
Basrah, Iraq.
ABSTRACT
The host parasite relationship of Schistosoma haematobium and the Balb/C mice are described.
Mice were exposed individually to 300 cercariae and worms were recovered, 8, 10, 12, 14, 16,
and 20-weeks post infection. The mean number of adult worms/mouse ranged between 15 and
40. The sex ratio of worms was always in favour of males, 8:1 at 10 weeks after infection, then it
decreased regularly to 3:1 at 20 weeks. From the 12th
and 14th
weeks post infection, a high
number of eggs was found in the liver and intestine respectively. At 14 weeks, eggs were also
found in spleen, lung, and kidney. Viable eggs were isolated from the liver of infected mice and
living miracidia were obtained. Faecal eggs were first observed in some mice by 16 weeks post
infection. The egg-induced pathology is described in a separate paper (1)
.
‫الخالصة‬
‫درست‬
‫العالقة‬
‫بين‬
‫طفيلي‬
‫المنشقة‬
‫البولية‬
‫والمضيف‬
‫باستخدام‬
‫فئران‬
‫بالب‬
/
‫س‬
.
‫حيث‬
‫عرض‬
‫كل‬
‫فئر‬
‫الى‬
300
‫سركاريا‬
‫وتم‬
‫عزل‬
‫الديد‬
‫ان‬
‫البالغة‬
‫بعد‬
8
،
10
،
12
،
14
،
16
،
‫وعشرين‬
‫اسبوعا‬
‫من‬
‫االصابة‬
.
‫كان‬
‫معدل‬
‫االصابة‬
15
-
40
‫طفيلي‬
/
‫فئر‬
‫وكانت‬
‫نسبة‬
‫الذكور‬
‫الى‬
‫االناث‬
1:8
‫بعد‬
‫ثمان‬
‫اسابيع‬
‫تدنت‬
‫الى‬
1:3
‫بعد‬
20
‫اسبوعا‬
.
‫وجدت‬
‫اعداد‬
‫كبيرة‬
‫من‬
‫البيوض‬
‫في‬
‫الكبد‬
‫واالمعاء‬
‫بعد‬
12
-
14
‫اسبوع‬
‫من‬
‫االصابة‬
,
‫بعض‬
‫من‬
‫هذه‬
‫البيوض‬
‫كانت‬
‫تحوي‬
‫على‬
‫مراسيديا‬
‫حية‬
.
INTRODUCTION
Very few studies have been carried out on S. haematobium mainly because of the difficulties in
maintaining its life cycle under laboratory conditions. The most widely used hosts on which
several generations of the parasite can be maintained are the hamster and the jird (Merione
Unguiculatus). The mouse, is a choice host for experimental research and for the maintenance of
cycles for most species of Schistosoma.

2. MJBU, Vol 13. No. 1&2, 1995
22
The mouse however, has been regarded as an unsuitable host for S. haematobium because of the
low percentage of adult worm development (2,3,4)
. Moreover, in all studies, attempts to infect
snails with miracidia hatched from eggs produced in mice always failed.
In this study the suitability of the mouse as a laboratory host for S. haematobium and the
distribution of parasite eggs in the mice tissues have been investigated.
MATERIALS AND METHODS
Bulinus truncatus (2-5 mm) in length which were maintained in our laboratory were individually
exposed to 5 miracidia of S. haematobium obtained from heavily infected school children from
Dhiqur governorate. Snails were maintained in glass containers and the cultures kept in
controlled temperature at 270
C in a 12hour light/dark cycle and fed on lettuce leaves.
Balb/C mice 6-8 weeks old were exposed individually to 300 freshly emerged cercariae using
padding technique (5)
. Starting from 8 weeks post infection mice were killed chloroform. The
abdominal and thoracic cavities were opened and worms collected from the hepatic portal blood
system and intestinal veins using the perfusion technique (6)
.
Six groups of 5 infected mice each were perfused at intervals of 8, 10, 12, 14, 16 and 20 weeks
after infection. Following perfusion, the entire spleen, lungs, kidneys, liver and the intestine were
removed and digested in 4% KOH. Five aliquot samples of 1 ml were taken with a pipette from
each of the 50 ml digest suspension and the number of eggs in each tissue calculated from their
mean count. Tissue culture chamber was used for this purpose. Faecal eggs were investigated for
each mouse on the day of perfusion.
The activity of miracidia originated from liver eggs were also checked.
Statistical analysis were based on log transformation of number of worms recovered from mice
and geometric means were calculated rather than the arithmetic means to reduce variability
which is inherent in such type of data. t-test was carried out on the log transformed data to
investigate differences in number of worms recovered after different periods of infection.
RESULTS
Table 1. Shows the worms recovered in the six groups of mice, perfused from 8 to 20 weeks post
infection. The geometric mean number of adult worms/mouse ranged from 13.9 at 20 weeks to
31.8 at 10 weeks post infection. The geometric means number of worms recovered after 16 and
20 weeks of infection were significantly lower than after 12 weeks of infection (t=4.5, 5.2, p
(0.05), while these means were not found to be significantly lower than those after 8 or 18 weeks
of infection.
The data show that there are marked variation in the sex proportion and worm returns from mice
at different time intervals. The mean sex rations of adult worms recovered in each group appears

3. MJBU, Vol 13. No. 1&2, 1995
23
to be in favour of males. Highest sex ratio 8:1 was noted at 10 weeks after infection and the
lowest 3:1 was noted at 14 weeks post infection. The overall sex ratio was 4.7 males to each
female.
Table 2 & 3 show the distribution of parasite eggs in mice tissues. The results also show that
after 8, 10 weeks post infection the eggs of the parasites were seen in the liver and intestine of
mice. At 14 and 16 weeks all mice had eggs in their liver, intestine, lung, spleen and, kidney. The
majority of the parasite eggs were retained in the liver first followed by intestine, lung, spleen,
and kidney.
Eggs isolated from the liver were hatched and active miracidia were checked. Attempts to infect
snails with miracidia hatched from eggs were in progress.
No eggs were found in mice urine at any time during this study.

4. MJBU, Vol 13. No. 1&2, 1995
24
DISCUSSION
The mouse has generally been regarded as a non-permissive host for S. haematobium largely on
the poor susceptibility to infection (7)
. In this study the maximum mean adult worms/mouse was
about 40. The mean worm return or the percentage of infecting cercariae that are recoverable as
mature worms has generally been recorded as 10% (8,4)
.
That ratio of male to female which is regarded as a measure of compatibility with host always in
favour of the males, a result which agrees with the finding of Kunts et al (10)
and Erickson et al
(11)
. The sex ratio of schistomoes in the vertebrate host can vary for a number of reasons (9)
. 0ur
results show a decrease in the ratio of male to female during the course of infection.
In this study, the first eggs appeared in the host tissues at 8 weeks post infection, mainly in the
liver and intestine. Eggs were found in the lungs, spleen and kidneys at 14 weeks post infection.
Smith et al (12)
showed that worms from hamster first produced eggs at 9.5 weeks. In our study
eggs were not found in the faeces, until 14 weeks and we never seen eggs in the urine. The
reduction in the mean number of adult worms/mouse was observed to terminate at 20-week post
infections.
Worms appear to be as long lived in mice and hamsters. Moore & Meleny (2)
and Kuntz &
Malakatis (10)
reported infections in mice and hamsters living in excess of 1 year. In man there
are records of infection persisting for up to 30 years (4)
.
Finally, we can conclude that the mouse S. haematobim combination could be used as a useful
model for further studies like infection of the snails with miracidia hatched from the mice liver
which is in progress now in our laboratory.

5. MJBU, Vol 13. No. 1&2, 1995
25
REFERNCES
1. Awad, A H H, Al-Yassin, T. M and Al-Khazzar, A. R. Histopathological studies of
schistosoma haematobium maintained in laboratory mice. Med. J. Bas. Univ. 1993 (in press).
2. Moore, D V, and Meleney, A I. Comparative susceptibility of common laboratory animals to
experimental infection with Schistosoma haematobium. J. Parasit. 1994; 40:392-397.
3. Nelson, G S, Amin. M A, Saoud, M F. A. and Teesdale, C. Studies on heterologous immunity
in Schistosomiasis I. Heterologous Schistosome immunity in mice. Bull. Wld. Hlth. org. 1968;
38: 9-17.
4. Cheever, A W, Duvall, R H and Hallack, T A. Hepatic fibrosis in Schistosoma haematobium
infected mice. Trans. Roy. Soc. Trop. Med Hyg. 1983; 77: 673-679.
5. Moore, D V, Yolles, T K and Meleney, H E. A comparison of common laboratory animals as
experimental hosts for Schistosoma mansoni. J. Parasit. 1949; 35: 156-170.
6. Smithers, Rand Terry, R T. The infection of laboratory hosts with cercaiae of of Schistosoma
mansoni and the recovery of adult worms. Parasit. 1965; 55: 695-700.
7. Smithers, S R. and Doenhoff, M J. Schistosomiasis. In: Immunology of parasitic infections.
2nd
ed. (ed.S. Cohen and K. S. Warren) 1982. PP.527-607. Oxford Blakwell scientific
Publications.
8. Taylor, M G and Andrews, B J. Comparison of the infectivity and pathogenicity of six species
of African Schistosomes and their hybrids. I. Mice and hamster. J. Hel. 1973; 47: 349-453.
9. Mitchell, G F, Garcia, F G, Wood. S M, Diasanta R Almonte A R, Calica, E Devern, K M and
Tiu, W V. Studies on the sex ratio of worms in Schistosoma infection. Parasit. 1990; 101. 27-34.
10. Kuntz, R E, and Malakatis G M. Susceptibility studies in Schistosomiasis III. Infection of
experimental hosts with Schistosoma haematobium in Egypt. Exp. Parasit. 1955; 4: 1-20.
11. Erickson, D G, Sadun, E H Lucia, H L, Von lichtenberg, F and Cheever, A W. Schistosoma
haematobium: Infection in five species. Exp. Parasit. 1971; 29: 126-137.
12. Smith, M A, Clegg, J A and Webbe, G. Culture of Schistosoma haematobium in vivo and in
vitro. Ann. Trop. Med. Parasit. 1976;
70:101-107.

6. MJBU, No- 1&2, 1995
vot 13,
Sabeeh lt AL-Mayah, MSc,^ Assistant
versj.ty of Basrah, Basrah lraq.
Lecturer,.Ciueg" df,1eao..tlon, Uni-
rir t )iSt
'| r,' , .1
. .r._ . .)
.- 1,.
DYNAI'IICAT ASPECTS OF SCHI$TOSOI'IA HAEI'{ATOBiUI'{
INI'ECTI0N :''AS E)(PERII|ENTAT !,l0DEt"
Abdul- llussein l{ Awad, Alim tr llYacoub, Sabeeh IIAl.trlayah
AbduL- {ussein H Awad, PhD; Assistant ProiesaoE
_College o! Eilucation-
University of Basrah, Iraq.'
Alim A- H Yacoub. PhD, mECM, Assistant ?rofsssor CoLtege of llledicine,
University of Ba3rah, Basrah.':Iraq.
ABSTRACT
The ho.st paraslte relationship of 99[!9!9!!4! haer€tobiur anal the Balb/C
nice are describeal- Mice lrere exposed individually to 300 cclcariae and
hrorns were recovereal, &t IO. L2, 14, 16, and 2o.t'eeks post lnfection-
The mean number of aalult !,qorms/nouse ranged between 15 and 40. The sex
ratio of worms was always in favour of males, 8:1 at 10 neeks after
infection, then it decreased regularly to 3:1 at 20 Feeks. Froh the 12th
anal 14th irees post infection. a high nuhber'of eggs ltas found in the
liver anal iniestine respectively. At 14 weeks. eggs r,tere also founal in
spl.een, lung and kidlney. viable eggs were Lsolateal from the llver of
infecteal mice;rand living diracialla l.rere obtaineal. Faecal eggs were first
observetl in lome mice by 16 lteeks post infection. the egg-Lndluced pa-
ttrologry is describeal in a separate papeE'_'.
INTRODUCTION.
1'
Very few studies have been carried, out on
S. haematobium mainly because of the dif*icul-
..---:----------.
ties in naintaining its life cycle under labora-
tory conditions. The most widely used hosts on
which several generations of the parasite can be
maintained are the hamster and the jirit
(Merione Unguicufatus). The mouse, is a choice
host for experimental research and for the

7. maintenance of cycles for most species of
Schi sto soma
The ,mouse however,
-has been regarded as an
unsuitable host for.s. haematobiui -u"I".r""
of
iii"."d:X.(2.rrr*t;::;; iF-"*"Hf
attempts to infect snails with rntracidi" t"t"h"d
from eggs produced in mice
"I""t;-;;;i;;_
In. this study the suitability of the mouse
laboratory host for .g. t
""r"tot-I.rr-'arra
distribution of parasLte eggs in the mLce
sues have been invesSigatedl-
MATERIALS AND METHODS
asa
the
tis-
EY+!+g: tEungatus (?-_s t*l in tength which were
maintained in our laboratory werE indiviauaiiy
exposed to 5 miracidia of G. 4gg,*"i"Uirr. ob_
tained from heavily infectect- ;ffifraIl"
f:"^_, llle": governorate. Snaits were- rnaintinea
r-n gtass containers and the cultures kept i;
controlled temDerature _ at ZZoc -ii-
a 12hour
light/dark
"y"-I" and fed
"rr-
i"tirr""' t""rr"..
Balb,/c mice 6-8 weeks old ,"""
""p".JJ-irrai",iarr_
aIly to 300 fresb+y emerged clrcariae using
padding technique( 5 ). starting f;;;-;"eks post_
infection mice were killed-
"tri".Jr".*. The
abdominal and thoracic cavities
""."-ii"""O and
$rorms collected from the hepatic p".iii. blood
system anf"{ntestinal veins rrsirrg ih"--perfusion
techniqug(o).
Six groups of 5 infected mice each,were perfused
at. intervals of B, LO,.]-,2, L4, i6
".a-ZO weeks
after infection. Follori.,g peri rsi""--in" entire
spreen, lungs, kidneys, riv?rr a"a-ihe intestine
$rere removed and digested in 4t XOff . five ali_
quot samples of 1 m1-were taken with i pipette
from each of the 5O nI- digest
"""p"""il and the
l:.P". of eggs in each tlssue
"li""iJi"a from
their mean count. Tissue
""ft"."--fi]*u"" $ras
used for this purpose. Faecal .gg= ,.;"-'investi_
.la
LL

8. gated for each mouse on the day of perfusion.
The activity of miracidia originated from liver
eggs rdere also checked-
The activity of miracidia originated from
were also checked -
liver
Statistical analysis were based on log transf,or-
mation of number of $rorms recovered, from mice
and geometric means were calculated rather than
the arithmetic means to reduce variability which
is inherent in such type of data. t-test was
carried out on the log transformed data to
investigate differences in number of worms
recovered after different periods of infection.
RESULTS
Table 1. Shows the worms recovered in the six
groups of mice, perfused from I to 20 weeks post
infection. The geometric mean nunber of adult
worms/mouse ranged from 13.9 at 20 weeks to 31.8
at 10 weeks post infection. The geometric means
nurnber of worms recovered. after 16 and 2O weeks
of infection were significantly lower than after
L2 weeks of infection (t=4.5, 5.2, p(0.05),
while these means were not found. to be signifi-
cantly lower than those after 8 or lSrweeks of
infection.
Ttre data show that there are marked variation in
the sex proportion and worm returns from mice at
different time intervals. The mean sex rations
of adult worms recovered in each group appears
to be in favour of males. I{ighesti sex ratio 8:1
r.ras noted at 10 weeks after infection and the
lowest,3:1 was noted at 14 weeks post infection.
The overall sex ratio was 4.7 males to each
female .
Table 2 & 3 show the distribution of parasite
eggs in mice tissues. The results also show that
after 8, 10 weeks post infection the eggs of the
parasites $rere seen in the liver and intestine
23

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12. of mice- At 14 and 16 weeks all mice had eggs in
their liver intestine, Iung, spleen and, kidney.
The maj ority of the Paraslte eggs were retained
in the liver first followed by intestine, 1ung,
spleen, and kidneY.
Eggs isolated from the lLver were hatched and
aiiive nriracidla were checked. Attenpts to
infect snails with niracidia hatched from eggs
wefe in Progress.
No eggs were found Ln nice urlne at any tine
durlng this study.
DISCUSSION
The mouse has generally been regarded as a non
permissive host for S. haematobium ,]qrgely on
th" poo" susceptibility to Lnfectionr'/. rn this
study the maximum mean adult worms/lnouse was
about 40- The mean worm return or the percentage
of infecting cercariae that are rrecoverable as
ifiltE?a rl"'^' has generally been recorded as
i
That ratio of mate to female which is: regarded
as a measure of conrpatibility with host always
in favour of the males, a result^.whi.ch agrees
^s t rt-.r-6 ^+ - r ( Iu J rnrt F*i aL<an
with t!9. finding of Kunts et al
^{. -t(rU Tha sct ratio of sc
and Erickson -
et al The sex ratio of schistomoes in the
host can vary for a nrimber of rea-
our results show a d.ecrease in the
vert?b{ate
sons -'.
ratio of male to female during the course of
infection. i
In this study, the first eggs appeared in the
host tissues at I eJeeks post infection, mainly
in the liver and intestine. Eggs were found in
the lungs, spleen and kiS4gYs at 14 weeks post
infection. Smith 9t aI'r'' 'r showed that $,orms
from hamster first Fiocluced eggs at 9-5 weeks.
In our study eggs were not found in the faeces,
until 14 weeks and we never seen eggs in the
urine. The reduction ln the mean number of adult

13. hrorms/mouse bras observed to terminate at 20 $reek
post infections.
Worms appear to be as lggg lived in mice and
hamslefs. Moore & Meleny o/ and Kuntz & Malaka-
1i5(ru) reported infections in mice dnd hamsters
living in excess of 1 year. In man there are
records of infection persisting for up to 30
years (4) -
Finally we can conclude that the mouse
S. haematobim combLnation could be used as a
usefuf rnoael for further studies like infection
of the snails with miracidLa hatched from the
mice liver which is "progress now in our labora-
tory,
REFERNCES
1. AvJad, A H H, Al-Yassin, T. M and A1-Khazzar,
A. R. Histopathological studies of schist
osoma haematobium naintained in labratory
mice. Med. J.'Bas- Univ- 1993 (in press).
2. Moore, D V, ahd Me1eney, A I. Compartive
susceptibilty, of conmon labortory animls to
experimental infection rdith Schistosoma
haematobium. .J. Parasit. L994, 4Oz.392-397.
I,
3. Nelson, G S, Amin. M A, Saoud, M F. A. and
Teesdale, C. 'Studies on heterologus immunity
in Schistosomj.asis I. Heterol-ogous Schisto-
some immunity in mice. Bull. $fld. H1th. org.
1968;38:9-17.
4. Cheever, A W, Duval1, R H and Hallack, T A.
Hepatic fibrosis in Schi stosomahaematobium
iniected mice. Trans. ffi
Hyg. 1983; ?7: 673-679.
5. Moore, D V, Yo1les, T K and Meleney, H E. A
comparison of common laboratory animals as
experimentatr. hosts for Schistosomamansoni . J.
Parasit- 1949; 35: 156-170.

14. 6. Smithers, Rand Terry, R T. The infection of
Iaboratory hosts with cercaiae of
Sctristosomamangqni and the recovery of adult
rornrs. P-;sit - 1965; 55: 695-700.
7- Smithers, S R. and Doenhoff, M J. Schistoso-
miasis. In Inununology of parasitic infec-
tions. 2nd ed. (ed. S. Cohen and K. S. War-
ren) 1982, PP.527-607. Oxford Blakwell Scien-
tific Publications.
8. Taylor, M G and Andrews, B J. Comparlson of
the infectivity and pathogenicity of six spe-
cies of African Schistosomes and their hy-
brids. I. Mice and hamstesr. J. Hel. 1973:.
47,349-453.
9. Mitchell , G F, Garcia, I'G, Wood. S M, Dias-
antaRA1monteAR, Ca1ica, E Devern, KMand
Tiu, W V. Studies on the sex ratio of worms
in Schistosoma infection. Parasit. 1990; 101:
27-34.
10. Kuntz, R E, and Malakatis G M. Suceptibiltiy
studies in Schistosomiasis III. Infection of
experimental hosts with Schistosoma
halnatobium in Egypt. fxp- earasi.t. 1955t 4:
L-20.
t_L. Erickson, D G, Sadun, E H Lucia, H L, Von
Lichtenberg, F and Cheever, A t{. Schistosoma
haematobium: Infection in five species. Exp.
Erasit- 197I; 29: L26-L37 .
12. smith, M A, Clegg, J A anil webbe, c. culture
of Schistosoma haernatobium in vivo and in
vitro. Ann. Trop. Med- Parasit. L976,
7 O:lOI-tO7 .
29

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