Surgical Science, 2012, 3, 452-456
http://dx.doi.org/10.4236/ss.2012.39089 Published Online September 2012 (http://www.SciRP.org/journal/ss)
Sumac (Rhus coriaria L.): Scolicidal Activity on Hydatid
Cyst Protoscoli ces
Mohammad Moazeni*, Maryam Mohseni
Department of Pathobiology, School of Veterinary Medicine, Shiraz University, Shiraz, Iran
Received May 4, 2012; revised June 10, 2012; accepted June 30, 2012
ABSTRACT
Background: Few anthelmintics are available for the treatment of hydatid disease caused by the parasite Echinococcus
granulosus. The appearance of resistance to synthetic anthelmintics and the adverse side effects of chemical drugs,
stimulated the research of alternatives, such as medicinal plants. In the present study, the scolicidal effect of methanolic
extract of sumac (Rhus coriaria) was investigated. Methods: Protoscolices were aseptically collected from sheep livers
containing hydatid cysts. Three concentrations of sumac extract (10, 30 and 50 mg/mL) were used for 10, 20 and 30
min. Viability of protoscolices was confirmed by 0.1% eosin staining. Results: While the rate of dead protoscolices was
16.93% in the ontrol group, when protoscolices were exposed to sumac extract at the concentration of 10 mg/mL , the
rate of dead protoscolices increased to 94.13%, 97.67% and 100% after 10, 20 and 30 minutes, respectively. The mor-
tality rate of protoscolices increased to 98.89%, and 100% when they were exposed to 30 mg/mL concentration of su-
mac extract for 10 and 20 minutes respectiv ely. One hundred percent mortality rate was observed at concen tration of 50
mg/mL after 10 min of exposure. Conclusions: This in vitro study showed that methanolic extract of R. coriaria may
be considered as an effective natural scolicidal agent.
Keywords: Hydatid Cyst; Scolicidal; Methanolic Extract; Sumac; Rhus coriaria
1. Introduction
Many tapeworms alternate their developmental cycle be-
tween intestinal stages in one host and tissue stages in
another. Hydatid disease is the result of tissue invasion
with the intermediate stage of a dog tapeworm, Echino-
coccus granulosus. The adult stage is a largely innocu ous
small tapeworm of dogs and other canids. The invasive
intermediate stage (metacestode) takes the form of an
enlarging cyst primarily in the liver and lungs of domes-
tic and wild herd animals. These cysts may be found sin-
gly, in clusters, or in such numbers that they pack the
peritoneal cavity. The principal sources of morbidity are
pressure effects from cyst size (up to 48 liters), location
in a sensitive organ (brain, reproductive tract, bone), or
cyst rupture with subsequent anaphylaxis or dissemina-
tion of the infection. The disease can be found in any part
of the world where slaughtering practices allow dogs to
consume the organs of infected animals. The parasite can
then comple te its developmen tal cycle [1,2]. Ther e are c ur -
rently three treatment options for hydatid disease, sur-
gery, ultrasound-gu ided asp iration , an d chemoth erap y [3 ].
Each of these modalities has limitations d epend ing on the
specific case. Chemotherapy is the preferred treatment
where, surgeons are not available or the cysts are too
numerous, and in inoperable cases, chemotherapy is the
only option. Chemotherapy has also been used as an ad-
junct to surgery for prophylaxis against spillage of cyst
contents [3,4]. For treatment of human hydatid disease,
the best agent available is the benzimidazole albend azole.
Most studies indicate that the efficacy of albendazole as
measured by the disappearance of a cyst is generally less
than 30% under ideal circumstances. Altogether, 60% of
cysts show some response in the course of therapy, in-
cluding shrinkage in size or detachment of cyst compo-
nents from the wall. Albendazole must be taken daily for
4 to 6 weeks, and this course should be repeated an addi-
tional two or three times. The poor response of this in-
fection to most chemotherapeutic agents has made hy-
datidosis primarily a surgical dis eas e, an d thu s the ro le of
chemotherapy is for prophylaxis against spillage during
surgery, for the treatment of inoperable cases, or for use
in areas without adequate surgical facilities. There is clea rly
a need for drugs that are more effective and easier to ad-
minister [2]. Many efforts have been made to discover
new antimicrobial compounds from various kinds of
sources such as plants, animals and microorganisms. Re-
cently, herbal medicines have increasingly been used to
treat many diseases including several infections [5].
*Corresponding autho
r
.
C
opyright © 2012 SciRes. SS

M. MOAZENI, M. MOHSENI 453
Sumac (Rhus coriaria L., family Anacardiaceae) g rows
wild in the region extend ing from the Canary Island over
the Mediterranean coastlin e to Iran and Afghanistan . It is
native to the Mediterranean and the Southeastern Anato-
lian Region of Turkey. The name is derived from
“sumâqâ”, meaning red in Syriac. The spice, produced by
grinding the dried fruit with salt, is used as a condiment
and sprinkled over kebabs and grilled meat as well as
over salads that often accompany these dishes. It has a
sour taste (pH 2.5) which is derived from the citric and
malic acids found in its juice [6]. Sumac is widely used
in Turkey and the Middle East, the fruits are red colored
and contain one seed. However, this part of the plant is
typically consumed as sp ice after drying and gr inding [7].
Sumac has been known to possess dietary and medicinal
properties. In folk medicine, it is used for treatment of
indigestion, anorexia, diarrhea, hemorrhagia and hyper-
glycemia [8]. Antioxidant [8-14], antidiabetic [15], hypo
glycemic [16], an tifibrog enic [17] and antitumorig enic [ 18 ,
19] properties of Sumac have been previously addressed.
It also has been shown to be useful in the treatment of
osteoarthritis [20].
The present study was conducted to evaluate the in
vitro scolicidal effect of methanolic extract of Sumac
(Rhus coriaria) on the proto s col i ces of hydatid cysts.
2. Materials and Methods
2.1. Collection of Protoscolices
Hydatid cysts from livers of naturally infected sheep wer e
obtained from Shiraz abattoir in southern Iran. The hy-
datid fluid was aseptically transferred into glass cylind ers
and left to set for 30 min. The protoscolices settled do wn
at the bottom of the cylinders. The supernatant was re-
moved and the yielded protoscolices were washed three
times with normal saline. Viability was assessed by mus-
cular movements and 0.1% eosin staining test. The live
protoscolices were finally transferred into a dark con-
tainer containing normal saline solution and stored at 4˚C
for further use.
2.2. Preparation of Sumac Extract
Dried fruits of R. Coriaria (Sumac) were powdered me-
chanically using a commercial electrical blender. To ob-
tain the methanolic extract, 100 g of dry Sumac powder
was added to 400 mL of pure methanol and mixed gently
for 1 h using a magnetic stirrer. The obtained solution
was left at room temperature for 24 h. The solution was
stirred again and filtered and then the solvent was re-
moved by evaporation in a rotating evaporator. The re-
maining semisolid material was then freeze-dried. The
obtained residue (6.2 g) was placed into a sterile glass
container and stored at 4˚C for further use.
2.3. Scolicidal Assay
In this study, three concentrations of sumac extract (10,
30 and 50 mg/mL) were used for 10, 20 and 30 min. To
prepare the sumac extract solution at 10, 30 and 50
mg/mL concentrations, 0.1, 0.3 and 0.5 g of dried extract
was dissolved in 10 ml of distilled water, respectively.
Then 2.5 mL of each sumac solution was placed in test
tubes, to which a drop of protoscolex-rich sediment was
added. The contents of the tubes were gently mixed .The
tubes were then incubated at 37˚C for 10, 20 and 30 min.
At the end of each incubation time the upper phase was
carefully removed so as not to disturb the protoscolices.
One milliliter of 0.1% eosin stain was then added to the
remaining settled protoscolices and mixed gently. The
upper portion of the solution was discarded after 15 min
of incubation. The remaining pellet of protoscolices was
then smeared on a manually scaled glass slide, covered
with a cover glass (24 × 50 mm), and examined under a
light microscope. The percentages of dead protoscolices
were determined by counting a minimum of 500 proto-
scolices. Non treated protoscolices were considered as a
control group in each experiment. The experiments were
performed in triplicate.
2.4. Viability Test
In the present study, eosin stain with the concentration of
0.1% (1 g of eosin powder in 1000 mL distilled water)
was used to check the viability of the protoscolices [21].
Fifteen minutes after exposure to the stain, the proto-
scolices with no absorbed dye were considered potent iall y
viable (Figure 1), otherwise they were recorded as dead
(Figure 2).
2.5. Statistical Analysis
Differences between the test and control groups were
analyzed with Chi-square test. Statistical analysis was
Figure 1. Live protoscolices after staining with 0.1% eosin.

M. MOAZENI, M. MOHSENI
454
Figure 2. Dead protoscolices after exposure to methanolic
extract of Rhus coriaria and staining with 0.1% eosin.
performed with GraphPad InStat software. P values less
than 0.01 were conside red to be signi fi cant.
3. Results
The mortality rate of hydatid cyst protoscolices after ex-
posure to different concentrations of the methanolic ex-
tract of R. coriaria following various exposure times are
presented in Tables 1-3. Rhus coriaria showed high
scolicidal activity and its methano lic ex tract was found to
be effective against protoscolices at all three concentra-
tions tested. While the mortality rate of protoscolices was
16.93% in the ontrol group, when protoscolices were
exposed to the R. coriaria extract at concentration of 10
mg/mL, the mortality rate increased to 94.13%, 97.67%
and 100% after 10, 20 and 30 minutes, respectively. The
mortality rate of hydatid cyst protoscolices after exposure
to concentration of 30 mg/mL of R. coriaria extract was
98.89%, and 100% after 10 and 20 minutes respectively.
One hundred percent mortality rate was observed with R.
coriaria extract at concentration of 50 mg/mL after 10
min of exposure. The difference between the scolicidal
effect of R. coriaria extract was statistically highly sig-
nificant (P < 0.0001) for all three concentrations and at
various exposure times, comparing to the control group.
4. Discussion
Few chemotherapeutic agents are available for the medi-
cal management of hydatid disease caused by the parasite
Echinococcus granulosus [2]. The control of helminthosis,
and, generally of all parasitic diseases is usually made
with synthetic anthelmintics. Up to date, many chemical
scolicidal agents have been used for inactivation of the
hydatid cyst protoscolices. Many of these scolicidal agent s
may cause undesirable complications that limit their use.
For example adverse side effects has been reported for
20% hypertonic saline, 20% silver nitrate, 0.5% - 1%
Table 1. Scolicidal effect of Rhus coriaria extract at the con-
centration of 10 mg/mL following various exposure times.
Exposure
time (min)ExperimentsProtoscolices Dead protoscolices Mortality rate (%)
10 1 789 727 92.14
2 964 912 94.60
3 753 720 95.61
Total 2506 2359 94.13
20 1 866 845 97.57
2 1341 1215 98.06
3 800 777 97.12
Total 3007 2937 97.67
30 1 586 586 100
2 880 880 100
3 963 963 100
Total 2429 2429 100
Control 2480 420 16.93
Table 2. Scolicidal effect of Rhus coriaria extract at the con-
centration of 30 mg/mL following various exposure times.
Exposure
time (min)ExperimentsProtoscolices Dead protoscolices Mortality rate (%)
10 1 1038 1018 98.07
2 1053 1051 99.81
3 624 616 98.71
Total 2715 2685 98.89
20 1 675 675 100
2 1320 1320 100
3 752 752 100
Total 2747 2747 100
Control 2480 420 16.93
Table 3. Scolicidal effect of Rhus coriaria extract at the con-
centration of 50 mg/mL following various exposure times.
Exposure
time (min)ExperimentsProtoscolices Dead protoscolices Mortality rate (%)
10 1 506 506 100
2 873 873 100
3 637 637 100
Total 2050 2050 100
Control 2480 2480 16.93
cetrimide, ethyl alcohol, and 20 mg/mL albendazole sul-
foxide [22]. The appearance of resistance to synthetic
anthelmintics stimulated the research of alternatives,
such as medicinal plants [23]. According to circum-
stances and depending on their efficacy, naturally pro-
duced plant anthelmintics offer an alternative that can
overcome some of these problems and is both sustainable
and environmentally acceptable [24]. Among the most
promising advances in the field of drug development is
discovering new molecules or novel uses of the already
available compounds with known safety and witho ut any
side effects [25]. A number of studies describe the in-
hibitory effects of different herbs and spices and their
volatile components on a variety of microorganisms. For

M. MOAZENI, M. MOHSENI 455
instance, sumac has been shown to possess antimicrobial
[26,27], antibacterial [6,28-31], antiviral [32], antimalar-
ial [33] and antifungal [34] properties.
In the present study we investigated the potency of
methanolic extract of sumac (R. coriaria) on the proto-
scolices of hydatid cyst. The results of our study showed
that sumac extract has a high scolicidal activity at the
concentrations of 10, 30 and 50 mg/mL after 30, 20 and
10 min of application, respectively. In previous study, we
investigated the protoscolicidal activity of garlic (Allium
sativum). Methanolic extract of garlic had 100% scoli-
cidal effect at a concentration of 25 mg/mL after 60 min
of exposure [23]. In the present study we observed a
higher scolicidal effect (100%) with methanolic extract
of sumac at a lower concentration (10 mg/mL) and in a
shorter exposure time (30 min).
Very limited literature is available on the mechanism
for the antimicrobial activity of herbs and spices 6. As
for the compounds in sumac which may be responsible
for the antimicrobial activity, more than 120 volatile
constituents have been identified by using gas chroma-
tography and mass spectroscopy, of which terpenoids and
aliphatic compounds were found occurr ing more freq u e n t l y
in six different varieties of sumac. Main constituents of R.
coriaria are terpene hydrocarbons (i.e. α-pinene, β-cary-
ophyllene and cembrene), oxygenated terpenes (i.e. α-ter-
pineol, carvacrol and β-caryophyllene alcohol) as well as
farnesyl acetone, hexahydrofarnesyl acetone and aliphatic
aldehydes [35].
This in vitro study showed that methano lic ex tract of R.
coriaria is an effective scolicidal agent. To the best of
our knowledge, this is the first report that investigates the
scolicidal efficacy of sumac extract on the protoscolices
of hydatid cysts. The results of this study allowed us to
suggest that sumac (R. coriaria) is likely source of new
compounds that could be used as an effective scolicidal
agent. Further studies will be necessary to identify and
isolate these active compounds. The results of present
study open the possibility of more investigations of in
vivo scolicidal effect of this traditional medicine.
5. Acknowledgements
The authors received financial support from Shiraz Uni-
versity (grant No. 87-GR-VT-24). We acknowledge Mr.
A. Motabi Alavi for his kind assistance during this study.
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