Pharmaceutical Biology

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Antischistosomal effect of holothurin extracted

from some Egyptian sea cucumbers

Mohammed H. Mona, Nahla E. E. Omran, Merveet A. Mansoor & Zainab M.

El-Fakharany

To cite this article: Mohammed H. Mona, Nahla E. E. Omran, Merveet A. Mansoor & Zainab
M. El-Fakharany (2012) Antischistosomal effect of holothurin extracted from some Egyptian sea
cucumbers, Pharmaceutical Biology, 50:9, 1144-1150, DOI: 10.3109/13880209.2012.661741

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Pharmaceutical Biology, 2012; 50(9): 11441150
2012 Informa Healthcare USA, Inc.
ISSN 1388-0209 print/ISSN 1744-5116 online
DOI: 10.3109/13880209.2012.661741

research ARTICLE

Antischistosomal effect of holothurin extracted from some

Egyptian sea cucumbers
Mohammed H. Mona, Nahla E.E. Omran, Merveet A. Mansoor, and Zainab M. El-Fakharany

Zoology Department, Faculty of Science, Tanta University, Tanta, Egypt

Abstract
Context: Holothuria polii (H. polii) Linnaeus (Holothuriidae), Actinopyga mauritiana (A. mauritiana) Quoy & Gaimard
(Holothuriidae) and Bohadschia vitiensis(B. vitiensis) Semper (Holothuriidae) are sea cucumbers inhabiting the coasts
of Egypt. Their tegument and the cuvierian gland contained a substance called holothurin that was used in traditional
medicine. These three species are abundant in the Egyptian coast, however there are no reports about their efficacy
as antiparasitic agent.
Objective: The antischistosomal effect of the holothurin extracted from the three species of sea cucumber is
investigated.
Materials and methods: The ethanol extract was made from the tegument of both H. polii and A. mauritiana while
it was made from the cuvierian gland of B. vitiensis. The body wall (or cuvierian gland) of the sea cucumber was
blended with 95% ethanol in a volume = 4 tissue weight. Extraction was done at room temperature for one day
then filtered. The ethanol was removed by evaporation using Rotavapour (BCHI 461 water bath REIII) at 40C. Later
the aqueous residue was placed in a vacuum oven at 20C for about 48 h to remove water. The resulting dried mass
was then stored at 4C until use. The percentage yield and the LD50 were calculated for each extract. Each extract
was administered orally to Shistosoma mansoni infected mice in acute and chronic phases of infection. The dose of
one-tenth of LD50 of each extract was administrated to mice (5.4, 62.2, and 10 mg/kg body weight/mouse for H. polii
extract (HPE), A. mauritiana extract (AME), and cuvierian gland of B. vitiensis, respectively) for 24 h. The effects of each
extract on the worm burden and total egg count was studied. The effects of each extract on the worm tegument
using scanning electron microscope (SEM) were investigated in vivo and in vitro.
Results: The percentage yield of cuvierian gland extract (CGE) was higher (70%) than the tegument AME (33.4%) and HPE
(9.3%). The 24 h LD50 of investigated sea cucumber ethanol extracts were 54.46, 627, and 100 mg/kg body weight/mouse
for HPE, AME, and CGE. Oral administration of HPE caused decrease in male and female worm burden of 30-day infected
mice to reach 60 and 90%, respectively. HPE decreased the egg count significantly in those mice with 30-day (1.75 egg
counts/g tissue, p < 0.05) and 45-day (3.25 egg counts/g tissue, p < 0.05) infections. SEM studies of recovered worms
from treated mice with all extracts showed different tegumental changes like formation of blebs, wrinkling, formation
of numerous pores, and rupturing of some tubercles. These effects were more pronounced in those worms treated in
vitro represented by severe shrinkage of the tegument, deformation of spines, rupturing, and collapsing of tubercles.
Discussion and conclusion: Results support the hypothesis that holothurin is a promising antischistosomal agent.
Keywords: Antiparasitic, glycoside, tegument

Introduction
Sea cucumber is a worldwide echinoderm of the class
Holothuroidea, with an elongated body and leathery
skin, which is found on the sea floor. Sea cucumbers
have long been used as a source of traditional medicines
in Malaysia (Hassan et al., 1996). Modern research has
shown that sea cucumber extracts have wound heal-
ing effects (Hassan et al., 1996), besides, they contain
anticoagulant, antithrombotic compounds, cholesterol,
lipid reducing compounds (Zancan & Mourao, 2004),

Address for Correspondence: Nahla E. E. Omran, Medical Campus, El-Bahr Street, Faculty of Science, Tanta University, Tanta, Egypt.
E-mail: nahlaayo@hotmail.com
(Received 25 August 2011; revised 08 November 2011; accepted 10 January 2012)

1144

anticancer compounds (Hatakeyama etal., 2002; Kariya
et al., 2004), antiinflammatory (Yamonouchi, 1955;
Nigrelli, 1972; Idid et al., 2001) and an antifungal com-
pounds (Sedov etal., 1990; Omran, 2006). Yoshida etal.
(2007) found that a lectin from sea cucumber Cucumaria
echinata Von Marenzeller (Cucumariidae) impaired the
development of the malaria parasite when produced by
transgenic mosquitoes.
Yamonouchi (1955) was the first to coin the term
holothurin to signify the biologically active compound
produced by the body wall of holothuroid cucumbers.
This compound is found also in the cuvierian tubules
and the coelomic fluid. Holothurin was isolated from
Pacific, Indo Pacific, and Atlantic species belonging to
the Holothuriidae family and particularly from species
of Holothuria and Actinopyga genera (Levin etal., 1984).
The structure of the holothurin extracted from the two
latter species has been established by Kitagawa et al.
(1978, 1982). Holothurin formed from triterpene glyco-
sides possess a 12--hydroxy-9(11)-ene fragment in the
glycone and disaccharide or tetrasaccharide part with an
O-sulphate group (Figure 1A). This structure is restricted
to species of Holothuria and Actinopyga genera (Stonik,
1986), while holothurin extracted from Bohadschia spe-
cies contains glycosides without an O-sulphate group
and with holost-9-(11)-en-12--3-ol as a genuine
Figure 1.Structure of some glycosides from Holothurin family
(Stonik 1986): A, Holothurin structure from Holothuria and
Actinopyga genera. B, Bohadschiosides from genus Bohadschia.
2012 Informa Healthcare USA, Inc.
Antischistosomal effect of holothurin 1145
glycone (Figure 1B). This type of glycoside was named
bohadschioside (Stonik, 1986).
Schistosomiasis is a disease caused by an infection
with parasitic blood flukes of the genus Schistosoma. This
fluke is distributed across Africa, the Arabian Peninsula,
South America, and some locations in the Caribbean.
More than 2 billion people, about one third of the worlds
population, are infected with these parasitic helmint-
hes (WHO, 2005). Several schistosomocide drugs were
administered to treat schistosomiasis, but praziquantel
(PZQ) showed the successful results in the treatment
because of its effectiveness and insignificant side effects
(Adam etal., 2005). However, after 30 years of PZQ usage,
a decreased susceptibility to the drug and the emergence
of drug-resistant strains was reported (Ismail etal., 1999;
King etal., 2000; Botros etal., 2005; Riad etal., 2009).
The present study investigated the antiparasitic activity
of sea cucumber extract against Schistosoma mansoni (S.
mansoni) as a model of endemic parasite in Egypt. Three
species of sea cucumber were used Holothuria polii (H.
polii), Bohadschia vitiensis (B. vitiensis), and Actinopyga
mauritiana (A. mauritiana). The efficacy of the holothu-
rin was tested at the acute and the chronic phases of the
infection and estimated by monitoring the worm burden,
egg count, and the worm surface topography in vivo and
in vitro using scanning electron microscope (SEM).
Materials and methods
Experimental animals
Adult male albino mice, Mus musculus were purchased
from Theodor Bilharz Research Institute, Egypt. Mice
were weighing between 1922 g at the beginning of the
experiments. The mice were kept in the laboratory for
1 week before the experimental work and maintained
on a standard diet and water available ad libitum. The
temperature in the animal room was maintained at 23
2C with a relative humidity of 55 2%. Mice were kept
at regular light and dark cycle. The experimental proto-
col was approved by local ethics committee and animals
research.
Mice infection
Collection of S. mansoni cercariae and the infection of
mice were carried out according to Christensen et al.
(1984). The mice were individually exposed to about 100
10 cercariae by the tail immersion technique.
Praziquantel
This drug is a chemotherapeutic treatment of schis-
tosome infection. It was purchased from Alexandria
Company for drugs and chemicals (Alexandria, Egypt).
PZQ was administered at days 30 and 45 in one oral dose
of 685 g/kg (b.w.) (El-Bolkiny & Al-Sharkawi, 1997).
Collection of sea cucumbers
The sea cucumber H. polii was collected from the inter-
tidal region at Abu-Kir coast (Mediterranean Sea) during
1146 M. H. Mona etal.
July 2006, while B. vitiensis and A. mauritiana were col-
lected from the intertidal zone of Hurgada coast (Red
Sea) in May 2006.
Preparation of crude extract
The ethanol extract was made from the tegument of both
H. polii and A. mauritiana while crude extract was made
from the cuvierian gland of B. vitiensis. All extracts were
prepared according to Shimada (1969) with some modi-
fications. Briefly, the body wall (or cuvierian gland) of the
sea cucumber was blended with 95% ethanol in a volume
= 4 tissue weight. Extraction was done at room tempera-
ture for one day then filtered. The ethanol was removed by
evaporation using Rotary evaporator (BCHI 461 water
bath REIII) at 40C. Later the aqueous residue was placed
in a vacuum oven at 20C for about 48 h to remove water.
The resulting dried mass was then stored at 4C until use.
Determination of the percentage of yield of different
extracts
The percentage yield of each extract was calculated
according to the following formula:
Dry weight of the extract
Percentage yields = 100
Dry weight of the
tissue extracted
Determination of LD50 of different crude extracts
Series of concentrations of each extract that permit the
computation of LD50 were prepared. Each concentration
was made using pure clean water and the exposure time
was 24 h. Data analysis was carried out using Finny pro-
gram with reliability interval of 95% (Finny, 1971).
Experimental design
Fifty-five mice were divided into three main groups:
1. Control uninfected (n = 5)
2. Infected with S.mansoni for 30 days (n = 25)
3. Infected with S.mansoni for 45 days (n = 25).
The second and the third groups were further divided
into five subgroups (five mice per group) as follows:
a. Mice treated with 5.4 mg/kg b.w./mouse of H. polii
extract (HPE).
b. Mice treated with 62.2 mg/kg b.w./mouse of A. mau-
ritiana extract (AME).
c. Mice treated with 10 mg/kg b.w./mouse of cuvierian
gland extract (CGE).
d. Mice treated with PZQ 685mg/kg b.w./mouse in a
single oral dose.
e. Control infected mice.
Worm recovery
Worms were collected from control and treated mice
by perfusion method as described by Christensen etal.
(1984). The collected worms from treated mice were
 Pharmaceutical Biology
examined by SEM to assess the in vivo effect of different
extracts on their tegument, while those collected from
the control mice were subjected to in vitro study.
In vitro study of the effect of different extracts on
S. mansoni worm
Male and female worms were divided into three glass
watches; in each, 7 worms in 1 ml saline (9%). One of
each extract was added to each glass watch in doses the
same as used in vivo to show whether they was affected
by endogenous factors of the mice which may decrease
their effectiveness. The time taken by the worm untill
death was calculated in each group.
Liver egg count
Total egg count was assessed as described by Pellegrino
etal. (1962).
SEM studies
The in vivo and in vitro effect of different extracts on the
worms tegument was assessed using AMRAY 1200 Bor
JEOL-JSM 5300 D.E.M. Specimens were washed many
times vigorously by shaking in 9% saline, and then fixed
in cold 2.5% glutraldahyde in phosphate buffer at 4C
overnight. Specimens were postfixed using 1% osmium
tetraoxide solution in 0.1 M phosphate buffer for 13 h
at 4C. The specimens were then washed for three times
in phosphate buffer and dehydrated in an ascending
series of ethanol at room temperature. Specimens were
mounted on the metal stubs, coated with gold using Spi-
modul vac spatter coater and examined with AMRAY
1200 Bor JEOL-JSM 5300 D.E.M.
Statistics
The obtained results were statistically analyzed using
Students t-test (Knapp & Miller, 1992) to determine
the significant differences between treated and control
specimens.
Results
The yield from the tegument of H. polii and A. mauritiana
was 9.3 and 33.4%, respectively, from the cuvierian gland
of B. vitiensis it was 70%.
As shown the CGE showed the highest yield followed
by AME and HPE.
To investigate the suitable dose of all investigated
extracts, the LD50 was estimated for each extract as shown
in Figures 2, 3 and 4 and Table 1. The one-tenth the LD50 of
each extract was administered to mice for 24 h. For HPE
the dose used was 5.4 mg/kg b.w./mouse while those for
AME and cuvierian gland of B. vitiensis were 62.2 and 10
mg/kg body weight/mouse, respectively.
Oral administration of HPE to 30-day infected mice
caused a significant decrease in male and female worm
burden reached 60 and 90%, respectively (Figure 5).
This reduction of the worm burden was accompa-
nied with a significant decrease in the liver egg count

Figure 2. Effect of Holothuria polii extract on mouse mortality.
Figure 3. Effect of Actinopyga mauritiana extract on mouse mortality
Figure 4. Effect of cuvierian gland extract on mouse mortality.
Table 1. Lethal dose values after 24 h exposure time to Sea
Cucumber extracts.
Species LD50
Holothuria polii 54.46 mg/mouse
Bohadschia vitiensis 100 mg/mouse
actinopyga Mauritiana 627.1 mg/mouse
(Figure 6).Administration of HPE to 45-day infected
mice caused insignificant decrease in the worm burden
(Figure 7). But it caused a significant reduction in the
liver egg count (Figure 8). Both AME and CGE caused
no significant changes on the worm burden and the egg
count of day 30 and day 45 infected mice (Figures 58).
Scanning electron microscopy of male worms revealed
that the tegument of the worm is smooth at the first third of
the body while its remaining parts were sponge like and cov-
ered with tubercles armed with spines (Figure 9A and 9B).
In vivo study of the effect of different extracts
(Figure9C, 9D, 9E and 9F) showed that all extracts had
destructive effect on the tegument including rupturing of
tubercles, folding of the tegument, formation of several
pores, and blebs. These effects were more pronounced
in those worms treated in vitro represented by severe
2012 Informa Healthcare USA, Inc.
Antischistosomal effect of holothurin 1147
Figure 5.The effect of different sea cucumber extracts on the
worm burden of 30-days infected mice. *Significant p < 0.05. HPE,
Holothuria polii extract; AME, Actinopyga mauritiana extract;
CGE, cuvierian gland extract; PZQ, praziquantel; cont 30, Control
infected mice for 30 days.
Figure 6. The effect of different extracts on the egg count of 30-days
infected mice. *Significant p < 0.05. HPE, Holothuria polii extract;
AME, Actinopyga mauritiana extract; CGE, cuvierian gland extract;
PZQ, praziquantel; cont 45, Control infected mice for 45 days.
Figure 7. The effect of different sea cucumber extracts on the worm
burden of 45-days infected mice. HPE, Holothuria polii extract;
AME, Actinopyga mauritiana extract; CGE, cuvierian gland extract;
PZQ, praziquantel; cont 30. Control infected mice for 30 days.
Figure 8. The effect of different extracts on the egg count of 45-days
infected mice. *Significant p < 0.05. HPE, Holothuria polii extract;
AME, Actinopyga mauritiana extract; CGE, cuvierian gland extract;
PZQ, praziquantel; cont 45, Control infected mice for 45 days.
shrinkage of the tegument, deformation of spines, rup-
turing and collapsing of tubercles (Figure 9G, 9H, 9I and
9J). In vitro treatment with AME caused immediate death
of the worm. While those treated with HPE and CGE died
after 20 min and 30 min, respectively.
1148 M. H. Mona etal.
Figure 9. Photomicrographs of control and treated Schistosoma
mansoni tegument. A, Control male worm showing os, vs and gpc.
B, The tegument of the middle part of the control worm covered
with Tb armed with s and sponge like Tg. C, In vivo treated worms
with HPE showing a few blebs (B) and transverse folds (F). D,
In vivo treated worm with HPE showing the RTg and deformed
Tb. E, In vivo treated male worms with AME showing transverse
folds (F) and (RTb). F, In vivo treated worms with CGE showing
the tegument with numerous blebs (B) and rough tegument with
transverse folds (F). G, In vitro treated worm with AME showing
severe shrinkage of both Tg and vs. H, In vitro treated worm with
AME showing CTb with deformed s. I, In vitro treated worm with
CGE showing CTb and RTb. J, In vitro treated worm with HPE
showing RTb and CTb. os, oral sucker; vs, ventral sucker; gpc,
gynechophoric canal; Tb, tubercles; s, spines; Tg, tegument; HPE,
Holothuria polii extract; RTg, rough tegument; AME, Actinopyga
mauritiana extract; RTb, ruptured tubercles; CGE, cuvierian
gland extract; CTb, collapsed tubercles.
 Pharmaceutical Biology
Discussion
The emergence of drug-resistant schistosome strains has
brought great attention toward natural bioactive com-
pounds. Several studies have been done to evaluate natu-
ral substances of plant origin as antischistosomal agents
such as Curcuma longa (Rizk etal., 2000), myrrh (Massoud,
1998; Sheir etal., 2001), garlic (Riad etal., 2009), and ginger
(Al-Sharkawi etal., 2007; Mostafa etal., 2011). To the best
of our knowledge the current study is the first trial to use
this substance (holothurin) as an antischistosomal agent.
Among the investigated extracts, HPE showed
decreased male and female worm burden to 60 and 90%,
respectively, in 30-day infected mice. This reduction was
accompanied with liver egg count reduction that reached
56%. Comparing these results with those obtained from
PZQ treated mice revealed that HPE may be a promising
antischistosomal agent. Another point that should be
taken into consideration is that the effect of HPE is signif-
icant when administered at an early stage of infection (30
days of infection; the acute phase). Comparable findings
were obtained by Riad etal. (2007) who reported a sig-
nificant reduction in the egg load after treating infected
mice with aqueous garlic extract at acute phase.
Despite the significant effect of HPE during the early
stage of infection, it did not reveal any significant change in
the worm burden of 45 day-infected mice. It caused, how-
ever, 56% reduction in the liver egg count. Accordingly, it
can be suggested that the reduction in the egg count is not
due to the decrease in the worm burden but it might be
due to its ability to decrease worm fecundity.
Recovered worms from treated mice with all extracts
showed different tegumental changes like formation of
blebs, wrinkling, formation of numerous pores, and rup-
turing of some tubercles. These tegumental alterations
would impair the functioning of the defence system of
the worm, so that it could easily be attacked by the hosts
immune system (Xiao etal., 2000). Moreover, it results in
the inability of the worm to adhere to the walls of the host
blood vessels, causing the schistosome to be dislodged
and moved by blood stream from mesenteric veins to
the portal vein and intravenous hepatic capillaries and
become lodged in the liver which is followed by the death
and disintegration of the parasites (Mehlhorn etal., 1981;
Riad etal., 2009; Gnanasekar etal., 2009).
There is no available information about the effect of
holothurin on the tegument. But there are other related
studies about the effect of antischistosome drugs that
reported tubercular disruption and loss of tubercular
spines as a result of treatment (Amin & Mikhail, 1989;
Mostafa & Soliman 2002; Shaohong etal., 2006; Taha &
Soliman 2007; Riad etal., 2009).
Although the in vivo administration of AME and
CGE showed no effect on the worm burden or on the
egg count, the SEM revealed severe destruction of the
worm tegument as manifested by severe shrinkage of the
tegument, collapsing and rupturing of tubercles beside
deformation of spines, suggesting that these two extracts
express strong schistosomocide. These defects could

be attributed to the act of their metabolic pathways in
infected mice that suppressed the activity of the extracts
biological compounds or it may be attributed to the dose
regimen or to the period of treatment that resulted in
low serum concentration of the curative compound that
reach mesenteric and portal vessels.
Conclusion
The results obtained suggest that sea cucumbers H. polii
can be considered as a natural source for a new adjuvant
antiparasitic drug with conventional therapy such as PZQ.
Further study must be conducted to improve the effect of
the other investigated extracts in vivo, and to examine
their effects on the liver status of the infected mice.
Acknowledgments
We express our sincere thanks and appreciation to Prof.
Amal Khalil Eskander and Prof. Mohamed Labib Salem
for their kind revision of the manuscript.
Declaration of interest
The authors report no conflicts of interest.
References
Adam I, Elwasila E, Homeida M. (2005). Praziquantel for the treatment
of Schistosomiasis mansoni during pregnancy. Ann Trop Med
Parasitol, 99, 3740.
Al-Sharkawi IM, El-Shaikh KA, Tabl GA, Ali JA. (2007). The effect of
ginger on Schistosoma mansoni infected mice. Delta J Sci, 31,
110.
Amin AM, Mikhail EG. (1989). Schistosoma mansoni: Tegumental
surface alterations following oxamniquine treatment of infected
mice. J Egypt Soc Parasitol, 19, 815826.
Botros S, Sayed H, Amer N, El-Ghannam M, Bennett JL, Day TA.
(2005). Current status of sensitivity to praziquantel in a focus of
potential drug resistance in Egypt. Int J Parasitol, 35, 787791.
Christensen NO, Gotsche G, Frandsen F. (1984). Parasitological
techniques for use in routine laboratory maintenance of
schistosomes and for use in studies on the epidemiology of
human and bovine Schistosomiasis. WHO Collaborating Centre
for Applied Malacology: Danish Bilharziasis Laboratory, 40.
El-Bolkiny YE, Al-Sharkawi IM. (1997). Intolererance of indomethacin
by liver of mice with Schistosomiasis mansoni involvement. J Union
Arab Biol, 7(A) Zool, 227245.
Finny DJ. (1971). Estimation of the median effective dose. In: Probit
Analysis, 3rd ed. Cambridge University. Great Britain, 2049.
Gnanasekar M, Salunkhe AM, Mallia AK, He YX, Kalyanasundaram
R. (2009). Praziquantel affects the regulatory myosin light chain
of Schistosoma mansoni. Antimicrob Agents Chemother, 53,
10541060.
Hassan YN, Yeap KH, Shahimi MM. (1996). Effect of sea cucumber
water extraction on systemic anaphylactic reaction. Malaysian J
Med Sci, (Supp.) 3, 3738.
Hatakeyama T, Matsuo N, Shiba K, Nishinohara S, Yamasaki N,
Sugawara H, Aoyagi H. (2002). Amino acid sequence and
carbohydrate-binding analysis of the N-acetyl-d-galactosamine-
specific C-type lectin, CEL-I, from the Holothuroidea, Cucumaria
echinata. Biosci Biotechnol Biochem, 66, 157163.
Idid SZ, Jalaluddin DM, Ridzwan BH, Bukhori A, Nor Hazlinah S, Hoo
CC. (2001). Effect of two extracts from Sticopus badionotus Slenka
upon induced pleurisity in rat. Pakistan J Biol Sci, 4, 12911293.
2012 Informa Healthcare USA, Inc.
Antischistosomal effect of holothurin 1149
Ismail M, Botros S, Metwally A, William S, Farghally A, Tao LF, Day
TA, Bennett JL. (1999). Resistance to praziquantel: direct evidence
from Schistosoma mansoni isolated from Egyptian villagers. Am J
Trop Med Hyg, 60, 932935.
Kariya Y, Mulloy B, Imai K, Tominaga A, Kaneko T, Asari A, Suzuki
K, Masuda H, Kyogashima M, Ishii T. (2004). Isolation and
partial characterization of fucan sulfates from the body wall of
sea cucumber Stichopus japonicus and their ability to inhibit
osteoclastogenesis. Carbohydr Res, 339, 13391346.
King CH, Muchiri EM, Ouma JH. (2000). Evidence against
rapid emergence of praziquantel resistance in Schistosoma
haematobium, Kenya. Emerging Infect Dis, 6, 585594.
Kitagawa I, Yamanaka H, Kobayashi M, Nishino T, Yosioka I, Sugawara
T. (1978). Saponins and sapogenols XXVII. Revised structures of
holotoxin A and holotoxin B, two antifungal oligoglycosides from
the sea cucumber Stichopus japonicus Selenka. Chem Pharm Bull,
26, 37223731.
Kitagawa I, Kobayashi M, Kyogoku Y. (1982). Marine natural products.
IX. Structural elucidation of triterpenoidal oligoglycosides from
the Bahamean sea cucumber Actinopyga agassizi Selenka. Chem
Pharmacol Bull, 30, 20452050.
Knapp RG, Miller MC. (1992). Defining normality using the predictive
value method In: Cinical Epidemiology and Biostatistics, 1st
ed. National Medical Series (NMS), Egyptian edition. CMASS
publishing Co, 5360.
Levin VS, Kalinin VI, Stonik VA. (1984). Chemical characters and
taxonomic revision of holothurian Bohadschia graeffei (Semper) as
refer to elucidation of a new genus. Biol Morya, 3, 3338.
Massoud A, Salama O, Bennett JL. (1998). Therapeutic efficacy of new
antischistosomal drug, derived from myrrh, in active intestinal
schistosomiasis complicated with hepatosplenomegaly. In: Tada I,
Kojima S, Tsuji M, eds. Proceedings of the 9th International Congress
of Parasitology (ICOPA IX), Chiba, Japan. Bologna: Monduzzi
Editore, 619623.
Mehlhorn H, Becker B, Andrews P, Thomas H, Frenkel J. (1981).
In vivo & in vitro experiments on the effects of praziquantel on
Schistosoma mansoni. A light and electron microscopic study.
Arzneimittelforschung, 31, 544554.
Mostafa OM, Soliman MI. (2002). Experimental use of black-seed oil
against Schistosoma mansoni in albino mice: II. Surface topography
of adult worms. Egypt J Med Lab Sci, 11, 7985.
Mostafa OM, Eid RA, Adly MA. (2011). Antischistosomal activity of
ginger (Zingiber officinale) against Schistosoma mansoni harbored
in C57 mice. Parasitol Res, 109, 395403.
Nigrelli F. (1972). The effect of holothurin on fish and mice with
sarcoma-188. Zoologyica, 37, 8990.
Omran, NEE. (2006). Fungicidal and bactericidal effects of holothurin
extracted from four species of sea cucumber inhabiting
Mediterranean Sea and Red sea coast of Egypt. Proc 4th Int Con
Biol Sci (Zool), 16.
Pellegrino J, Oliveira CA, Faria J, Cunha AS. (1962). New approach to
the screening of drugs in experimental Schistosomiasis mansoni in
mice. Am J Trop Med Hyg, 11, 201215.
Riad NHA, Taha HA, Mahmoud YI. (2009). Effects of garlic on albino
mice experimentally infected with Schistosoma mansoni: A
parasitological and ultrastructural study. Trop Biomed, 26, 4050.
Riad NH, Fares NH, Mostafa OM, Mahmoud YI. (2007). The effect of
garlic on some parasitological parameters and on hepatic tissue
reactions in experimental Schistosomiasis mansoni. J App Sci Res,
3, 949960.
Rizk M, Hafez S, Farouk H. (2000). Measurement of urea cycle
enzyme activities in mice under the influence of different stages
of Schistosoma mansoni infection and Curcuma longa treatment.
J Egypt Ger Soc Zool, 32, 319333
Sedov AM, Apollonin AV, Sevastianova EK, Alekseeva IA, Batrakov SG,
Sakandelidze OG, Likhoded VG, Stonik VA, Avilov SA, Kupera EV.
(1990). [Stimulation of nonspecific antibacterial resistance of mice
to opportunistic Gram-negative microorganisms with triterpene
glycosides from Holothuroidea]. Antibiot Khimioter, 35, 2326.
1150 M. H. Mona etal.
Shaohong L, Kumagai T, Qinghua A, Xiaolan Y, Ohmae H, Yabu Y,
Siwen L, Liyong W, Maruyama H, Ohta N. (2006). Evaluation
of the anthelmintic effects of artesunate against experimental
Schistosoma mansoni infection in mice using different treatment
protocols. Parasitol Int, 55, 6368.
Sheir Z, Nasr AA, Massoud A, Salama O, Badra GA, El-Shennawy
H, Hassan N, Hammad SM. (2001). A safe, effective, herbal
antischistosomal therapy derived from myrrh. Am J Trop Med Hyg,
65, 700704.
Shimada S. (1969). Antifungal steroid glycoside from sea cucumber.
Sci, 83, 735737.
Stonik VA. (1986). Some terpenoid derivatives from echinoderms and
sponges. Pure Appl Chem, 58, 423436.
Taha HA, Soliman MI. (2007). Antischistosomal activity of
3-substituted-5-(2-aryl-2-oxoethyl)-2, 4-dioxo-1, 3-thiazolidine
(Ro-354). Inter J Agri Biol, 9, 8793.
 Pharmaceutical Biology
WHO. (2005). Deworming for health and development. In: Report
of the Third Global Meeting of the Partners for Parasite Control.
Geneva: World Health Organization, 51.
Xiao SH, Shen BG, Chollet J, Utzinger J, Tanner M. (2000). Tegumental
changes in 21-day-old Schistosoma mansoni harboured in mice
treated with artemether. Acta Trop, 75, 341348.
Yamonouchi T. (1955). On the poisonous substances contained
in holothurin. Publication of Seto Marine Biol Lab, 41,
183202.
Yoshida S, Shimada Y, Kondoh D, Kouzuma Y, Ghosh AK, Jacobs-
Lorena M, Sinden RE. (2007). Hemolytic C-type lectin CEL-III
from sea cucumber expressed in transgenic mosquitoes impairs
malaria parasite development. PLoS Pathog, 3, e192.
Zancan P, Mouro PA. (2004). Venous and arterial thrombosis
in rat models: Dissociation of the antithrombotic effects of
glycosaminoglycans. Blood Coagul Fibrinolysis, 15, 4554.