Experimental Parasitology 147 (2014) 715

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Experimental Parasitology
j o u r n a l h o m e p a g e : w w w. e l s e v i e r. c o m / l o c a t e / y e x p r

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A description of parasites from Iranian snakes

Vahid Nasiri a,b,*, Iraj Mobedi c, Abdolhossein Dalimi b, Abbas Zare Mirakabadi d,
Fatemeh Ghaffarifar b, Shohreh Teymurzadeh d, Gholamreza Karimi a, Amir Abdoli b,
Habibollah Paykari a
a

Department of Parasitology, Razi Vaccine and Serum Research Institute, Karaj, Alborz, Iran
Departments of Parasitology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
c Department of Parasitology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran
d
Department of Venomous Animals and Antivenom Production, Razi Vaccine and Serum Research Institute, Karaj, Alborz, Iran
b

H I G H L I G H T S

These are the rst data on the

parasitic fauna of Iranian terrestrial
snakes.
Little is known of the parasitic
fauna of terrestrial snakes in Iran.
This study demonstrates the high
prevalence of parasites among
Iranian snakes.

* Corresponding author. Fax: 009826-34552194.

E-mail address: v.nasiri@rvsri.ac.ir (V. Nasiri).
http://dx.doi.org/10.1016/j.exppara.2014.09.007
0014-4894/ 2014 Elsevier Inc. All rights reserved.

G R A P H I C A L

A B S T R A C T

V. Nasiri et al./Experimental Parasitology 147 (2014) 715

A R T I C L E

I N F O

Article history:
Received 23 March 2014
Received in revised form 19 September
2014
Accepted 24 September 2014
Available online 6 October 2014
Keywords:
Iranian snakes
Parasitic fauna
Intestinal parasites
Hemoparasites

A B S T R A C T

Little is known of the parasitic fauna of terrestrial snakes in Iran. This study aimed to evaluate the parasitic infection rates of snakes in Iran. A total of 87 snakes belonging to eight different species, that were
collected between May 2012 and September 2012 and died after the hold in captivity, under which they
were kept for taking poisons, were examined for the presence of gastrointestinal and blood parasites.
According to our study 12 different genera of endoparasites in 64 (73.56%) of 87 examined snakes were
determined. Forty one snakes (47.12%) had gastrointestinal parasites. In prepared blood smears, it was
found that in 23 (26.43%) of 87 examined snakes there are at least one hemoparasite. To our knowledge, these are the rst data on the internal parasitic fauna of Iranian terrestrial snakes and our ndings
show a higher prevalence of these organisms among them.
2014 Elsevier Inc. All rights reserved.

1. Introduction
More than 3500 species of snakes have been found around the
world, less than 10% of which are venomous (Bawaskar, 2004; Blackman and Dillon, 1992; Meenatchisundaram and Michael, 2009;
Warrel, 2005). In Iran, 69 species of snakes assigned to 37 genera
in six families have been identied, of which 36 species are nonvenomous, 25 species are venomous and 8 species are semivenomous (Dehghani, 2010; Lati, 2000; Zare Mirakabadi and
Teymurzadeh, 2008).
The class Reptilia, consisting of more than 6000 species, is host
to a wide variety of protozoan and metazoan parasites and virtually 100% of free-ranging reptiles harbor some kind of parasites and
however, the presence of these organisms is not necessarily associated with a disease state (Barnard and Upton, 1994; McFarlen, 1991).
Breeding snakes in captivity, particularly in a semi-extensive system,
is an activity that exposes the animals to the action of several parasites (Mader, 1996). Intestinal helminths, pentastomids, and intestinal
and blood protozoa are common endoparasites of captive or wild
reptiles that these animals may serve as denitive, intermediate,
accidental or paratenic hosts (Frye, 1991; Greiner and Mader, 2006;
Hernandez-divers, 2006; Jackson and Cooper, 1981; Macarthur et al.,
2004). The prevalence of parasites with a monoxenic cycle is higher
than that of heteroxenic-cycle parasites, since the latter require an
intermediate host to complete their cycle (Rey, 2001). The transmission of monoxenic-cycle parasites is favored by the physical and
organic conditions of the captivity, thus a snake can contaminate
other animals or even reinfect itself with its own feces. In addition,
the captivity conditions or the related stress may be responsible for
the acquisition or increase of the parasitic infection (Klingenberg,
1993). The result of this parasitism is competition with the host animal
for food, removal of tissue and uid, blocking of lymph and blood
vessels, edema, ulcerations, necrosis, and anemia (Barnard, 1983;
Frank, 1981; Frye, 1991; Marcus, 1981).

Although a considerable species of reptiles are present in Irans

ecosystems, there is not enough information about the parasitic fauna
of reptiles and their role in transmission of veterinary and zoonotic
disease (Yousse et al., 2013). In a research in Iran, a Natrix natrix
snake was found to be infected with the worm that belong to the
genus Ophiotaenia (Yousse et al., 2010). In a case report study, myiasis
due to Musca domestica was described in a Pseudocerastes persicus
snake and a lesion was found on its body where 14 live larvae of M.
domestica was removed (Dehghani et al., 2012). An investigation
showed that 1 turtle (Mauremys caspica caspica), 11 grass snakes
(Natrix natrix) and 5 dice snakes (Natrix tessellata) have been infected with Telorchis assula in Mazandaran, north of Iran (Yousse
et al., 2013). In another research, one European glass lizard, Pseudopus
apodus, and three European grass snakes, Natrix natrix, were examined for helminths and found that Pseudopus apodus harbored
one species of Nematoda, Entomelas entomelas and N. natrix harbored 1 species of Digenea, Telorchis assula, 1 species of Cestoda,
Ophiotaenia europaea, and 1 species of Nematoda, Rhabdias fuscovenosa
(Halajian et al., 2013). Recently it have been reported that 18 snakes,
including 9 N. natrix and 9 N. tessellata from Mazandaran Province,
north of Iran were infected with parasitic helminths including 1 Nematode: Rhabdias fuscovenosa (larva), 1 Digenea: Telorchis assula and
1 Cestoda: Ophiotaenia europaea (Yosse et al., 2014).
In this study the parasitic fauna of native Iranian snakes that were
taken from the wild and kept in captivity were investigated.
2. Materials and methods
A total of 87 snakes representing eight species that were collected between May 2012 and September 2012 from various
provinces of Iran sent to the department of Venomous Animals and
Antivenom Production, Razi Vaccine and Serum Research Institute. These parasites were kept under captivity and after ding
transferred immediately to the Parasitology laboratory of Razi

Table 1
The taxonomic characterization of examined snakes and number of their parasites.
Scientic name of snakes

Common name

Number of
examined
snakes

Number of
positive
snakes for
intestinal
parasitic

Number of
positive
snakes for
hemoparasites

Number of
positive
snakes for
any parasites

Pseudocerastes persicus eldi

Naja oxiana
Vipera albicornuta
Vipera lebetina obtusa
Vipera ursinii eriwanensis
Agkistrodon intermedius caucasicus
Natrix natrix
Coluber caspius gmelin
Total

Persian horned viper

Central Asian cobra
Zigzag mountain viper
West-Asian blunt-nosed viper
Transcaucasian meadow viper
Caucasian pit viper
European grass snake
Caspian whip snake

23
10
5
20
2
23
1
3
87

10
7
5
10
2
5
1
1
41

13
3
0
6
0
0
0
1
23

23
10
5
16
2
5
1
2
64

V. Nasiri et al./Experimental Parasitology 147 (2014) 715

Fig. 1. Acanthocephalan larval detected on the intestinal wall surface.

Fig. 2. The detected Telorchis assula from Natrix natrix.

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V. Nasiri et al./Experimental Parasitology 147 (2014) 715

Fig. 3. The detected (A) Ophiotaenia sp. and (B) one unknown species from Natrix natrix.

Fig. 4. The detected nematodes belong to superfamily Rhabditoidea, Strongyloides sp. from one Vipera ursinii eriwanensis and four Naja oxiana.

V. Nasiri et al./Experimental Parasitology 147 (2014) 715

11

Fig. 7. The detected Blastocystis sp. from one Vipera lebetina obtuse.

Fig. 5. The detected Nematode: Ascarididae from intestine of one Naja oxiana.

Vaccine and Serum Research Institute. The snakes body wall was
opened by a longitudinal incision and organ surfaces, mesenteries
of the body cavity, the lumen of the digestive tract and the lungs
were examined visually for parasites. Subsequently, the viscera were
placed in a Petri dish with normal physiological saline and examined for helminths under a dissecting microscope. The recovered
helminths were cleaned carefully and, xed in 70% alcohol and
stained with acetocarmine. The Protozoan parasites were identied by prepared direct wet mount smears and modied Ziehl
Neelsen staining of fecal samples and examination of them under
light microscope. The fecal samples were placed in vials with 2.5%
K2Cr2O7 and 10% formalin for examination of the fecal contents for
any parasite eggs or oocysts and sporulation of any oocysts found
in the samples.
For identication of hemoparasites, thin blood smears were prepared from cardiac blood of each snake and then air dried, xed
in absolute methanol, stained with Giemsa stain and examined under
light microscope.
Collected parasites were deposited in the Museum of Parasitology Department, Razi Vaccine and Serum Research Institute, Karaj,
Alborz, Iran.

Fig. 6. The detected Eimeria sp. from one Vipera ursinii eriwanensis and one Vipera
albicornuta.

Fig. 8. The detected Nyctotheroides sp. from one Pseudocerastes persicus eldi.

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V. Nasiri et al./Experimental Parasitology 147 (2014) 715

Fig. 9. The detected Hexamita batrachorum (Giemsa stain).

3. Results
3.1. Intestinal parasites
3.1.1. Intestinal worm infection
Twelve different genus of endoparasites were determined in 64
(73.56%) out of 87 examined snakes. In the majority of snakes, two

or more species of parasites were found (see details in Table 1). At

necropsy, an Acanthocephalan larval stage (Centrorhynchus
corvi) was found on the outer surface of the intestinal wall
of seven snakes (2 Vipera lebetina obtuse; 2 Vipera albicornuta; 2 Naja
oxiana and 1 Coluber caspius gmelin). Local necroses were observed on the intestinal wall surface of snakes with Centrorhynchus
corvi (Fig. 1).

Fig. 10. Intraerythrocytic gametocytes in Naja oxiana.

V. Nasiri et al./Experimental Parasitology 147 (2014) 715

13

Fig. 11. Intraerythrocytic hemoparasites in Pseudocerastes persicus eld. (AD) Intraerythrocytic gametocytes with different pattern of erythrocyte shape changes. (E) Developing schizonts in the liver of infected snake. (F) Unknown organism from liver.

An adult stage of one species of trematode (Digenea: Telorchiidae:

Telorchis assula) (Fig. 2), two species of cestoda (A: Ophiotaenia sp.
and B: one unknown species) (Fig. 3) were detected from the intestine of a Natrix natrix (European Grass Snake).
Adult stages of two genus of nematode were detected, including: one belonging to the superfamily Rhabditoidea (Strongyloides
sp.) from the intestine of ve snakes (one Vipera ursinii eriwanensis
and four Naja oxiana) (Fig. 4) and one that belongs to family
Ascarididae from the intestine of one Naja oxiana (Fig. 5).
3.1.2. Intestinal protozoa infection
Eimeria sp. was detected in two snakes (one Vipera ursinii
eriwanensis and one Vipera albicornuta) and in both snakes with
Eimeria sp. infection, petechia was observed in the mucous layer
of the intestinal tract (Fig. 6). A large number of Blastocystis sp. was

Fig. 12. Intraerythrocytic gametocytes in Coluber caspius gmelin.

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V. Nasiri et al./Experimental Parasitology 147 (2014) 715

Fig. 13. Intraerythrocytic gametocytes in Vipera lebetina obtusa.

detected in one snake (Vipera lebetina obtuse) (Fig. 7). Moreover, Ciliated protozoa (Nyctotheroides sp.) was detected in a snake
(Pseudocerastes persicus eldi) (Fig. 8).
A large numbers of Hexamita batrachorum (a agellate protozoa belonging to Phylum Sarcomastigophora, Class Mastigophora) was
detected in intestinal mucosa and stool of 27 snakes (9 Pseudocerastes
persicus eldi, 8 Vipera lebetina obtuse, 3 Agkistrodon intermedius
caucasicus, 1 Vipera ursinii eriwanensis, 2 Vipera albicornuta and 4
Naja oxiana) (Fig. 9). This parasite was the most frequent organism found in snakes (31.03%).
3.2. Hemoparasites
Hemoparasites were observed in 23 (26.43%) out of 87 examined snakes. Many of the infected red blood cells were distorted in
size and shape; however, some infected cells were similar to
uninfected erythrocytes. We could not to see sporogonic stages,
thus we could not determine the parasites genus. Microscopic examination of Giemsa stained slides showed infection of red blood
cells of 13 Pseudocerastes persicus eldi, 6 Vipera lebetina obtusa, 3
Naja oxiana and 1 Coluber caspius gmelin with Hemoparasites
(Figs. 1013).
4. Discussion
The reptiles have important impacts on different aspects of their
ecology and thus demands deep research regarding the inuence
of these animals and their ora on people and autochthonous animal
species. The varieties of different pathogens in these species are very
large. The presence of several pathogens in one host and stressful
situations can have a negative inuence on the health status. Furthermore, there is a possibility of pathogen transmission to human.
A healthy reptile has a number of pathogens, all kept in check by
a healthy immune system and the benecial gut ora. When a reptile
is highly stressed or under prolonged moderate to severe stress, the
immune system falters. In cases of improper environmental temperatures, starvation, or prolonged dehydration, the benecial gut
ora die off and organisms benign in small numbers gain ascendancy and start causing problems (Rataja et al., 2011).
It is widely known that under captivity the stress to which
animals are submitted to can induce the settlement of several pathologies, mainly those related to parasites (Leinz et al., 1989; Santos
et al., 2008; Siqueira et al., 2009) and because parasites can reach

high infection loads in hosts in captivity, can be very problematic

and may cause the death of hosts (Klingenberg, 1993). So for conservation efforts involving captive breeding, an understanding of the
parasites of a particular species is crucial. With this view, it is necessary to provide data on the parasitic fauna of Iranian snakes, in
order to permit the establishment of prophylactic procedures to
properly reduce the mortality of captive animals.
The present study was carried out to evaluate the parasitic infections in wild snakes. The study was proposed because we observed
that, in captivity, snakes presented a high prevalence (above 70%)
of infection by different parasite species. The present investigation showed that 64 (73.56%) of 87 analyzed snakes arrived for
captivity already infected by at least one parasite species. The results
from this study are important for the sanitary handling of these important animals during captivity. Other different species were also
detected that similar parasite were described in the previous literatures (Klingenberg, 2000; Mader, 1996; Telford, 2009). In two
snakes a wide dilatation and destruction of gastrointestinal tract
was seen and Modied ZiehlNeelsen staining was positive for
Eimeria sp. The Hemogregarines (Apicomplexa: Adeleorina) are,
intraerythrocytic protozoan parasites that infect a wide variety of
vertebrates (Davies and Johnston, 2000; Telford, 2009) and are considered common Hemoparasites of reptiles and this was also true
in our study.
In conclusion, to our knowledge, these are the rst data on the
parasitic fauna of Iranian terrestrial snakes and our ndings show
a higher prevalence of these organisms among them. Control and
prevention of many parasitic diseases is associated with breaking
the cycle of transmission, and there is no shadow of doubt that
precise information about diseases and their causative agents is the
major key to control of them and thus, further precise parasitological investigations are required due to the noticeable unexplored
area of our country in order to ascend our knowledge concerning
parasites of snakes and probable zoonotic and veterinary importance of them.
References
Barnard, S.M., 1983. A review of some fecal pseudoparasites of reptiles. J. Zoo. Anim.
Med. 14, 7988.
Barnard, S.M., Upton, S.J., 1994. A Veterinary Guide to the Parasites of Reptiles:
Protozoa. Krieger Publishing Co., Malabar, FL. 154 pp.
Bawaskar, H.S., 2004. Snake venoms and antivenoms: critical supply issues. J. Assoc.
Physicians. India 52, 311.

V. Nasiri et al./Experimental Parasitology 147 (2014) 715

Blackman, J.R., Dillon, S., 1992. Venomous snakebite: past, present, and future
treatment options. J. Am. Board Fam. Pract. 5 (4), 399405.
Davies, A.J., Johnston, M.R.L., 2000. The biology of some intraerythrocytic parasites
of shes, amphibia and reptiles. Adv. Parasitol. 45, 1107.
Dehghani, R., 2010. Animal Natural Toxins. Environmental Toxicology, rst ed. Tehran
Tak Derakhat and Kashan University of Medical Sciences, Iran. p. 389-45.
Dehghani, R., Sedaghat, M.M., Sabahi Bidgoli, M., 2012. Wound myiasis due to Musca
domestica (Diptera: Muscidae) in Persian horned viper, Pseudocerastes persicus
(Squamata: Viperidae). J. Arthropod Borne Dis. 6 (1), 8689.
Frank, W., 1981. Endoparasites. In: Cooper, J.E., Jackson, O.F. (Eds.), Diseases of the
Reptilia, vol. 1. Academic Press, London, pp. 291358.
Frye, F.L., 1991. Biomedical and Surgical Aspects of Captive Reptile Husbandry, second
ed. Krieger Publishing, Melbourne, FL.
Greiner, E.C., Mader, D.R., 2006. Parasitology. In: Mader, D.R. (Ed.), Reptile Medicine
and Surgery, second ed. Saunders Elsevier, St. Louis, MI, pp. 43364.
Halajian, A., Bursey, C.R., Goldberg, S.R., Gol, S.M.A., 2013. Helminth parasites of the
European glass lizard, Pseudopus apodus (Squamata: Anguidae), and European
grass snake, Natrix natrix (Serpentes: Colubridae), from Iran. Comp. Parasitol. 80,
151156.
Hernandez-divers, S.J., 2006. Reptile parasites summary table. In: Mader, D.R. (Ed.),
Reptile Medicine and Surgery, second ed. Saunders Elsevier, St. Louis, MI, pp.
11591170.
Jackson, O.F., Cooper, J.E., 1981. Cooper, J.E., Jackson, O.F. (Eds.), Diseases of the Reptilia,
vol. 1 & 2. Academic Press, New York, NY.
Klingenberg, R.J., 1993. Understanding Reptile Parasites: A Basic Manual for
Herpetoculturists & Veterinarians. Advanced Vivarium Systems, Lakeside, CA.
Klingenberg, R.J., 2000. Diagnosing parasites in old world Chameleons. Exotic DVM
1, 1721.
Lati, M., 2000. The Snakes of Iran, third ed. Environment Protection Organization,
Tehran.
Leinz, F.F., Janeiro-Ciquini, T.R.F., Ishizuka, M.M., 1989. Sobrevivncia de Bothrops
jararacussu (Serpentes, Viperidae, Crotalinae) mantidas em cativeiro. Mem. Inst.
Butantan 51, 3338.
Macarthur, S., Wilkinson, R., Meyer, J., 2004. Medicine and Surgery of Tortoises and
Turtles. Blackwell Publishing, Ames, IA.

15

Mader, D.R., 1996. Reptile Medicine and Surgery. WB Saunders, Philadelphia, PA.
Marcus, L.C., 1981. Veterinary Biology and Medicine of Captive Amphibians and
Reptiles. Lea & Febiget, Philadelphia, PA.
McFarlen, J., 1991. Commonly occurring reptilian intestinal parasites. In: Proceedings
of the American Association of Zoo Veterinarians. Calgary, Canada, pp. 120127.
Meenatchisundaram, S., Michael, A., 2009. Snake bite and therapeutic measures:
Indian scenario. Indian J. Sci. Tech. 2 (10), 6973.
Rataja, V., Lindtner-Knic, R., Vlahovic, K., Mavri, U., Dovc, A., 2011. Parasites in pet
reptiles. Acta Vet. Scand. 53, 33.
Rey, L., 2001. Parasitologia, third ed. Guanabara Koogan, Rio de Janeiro.
Santos, K.R., Takahira, R.K., Rall, V.L.M., Caldern, C., Sequeira, J.L., Silva, R.J., 2008.
Pulmonary, microbiological and hematological changes in Crotalus durissus
terricus (Serpentes, Viperidae) parasitized by nematodes of the genus Rhabdias
(Nematoda, Rhabdiasidae). Arq. Bras. Med. Vet. Zootec. 60, 667674.
Siqueira, L.R., Panizzutti, M.H.M., Muniz-Pereira, L.C., Pinto, R.M., 2009. Gross lesions
induced by nematodes of Bothrops jararaca and Bothrops alternatus in Brazil
with two records. Neotrop. Helminthol. 3, 2933.
Telford, S.R., 2009. Hemoparasites of the Reptilia: Color Atlas and Text. CRC Press,
Boca Raton, London, and New York.
Warrel, D.A., 2005. WHO Guidelines for the Clinical Management of Snake Bite in
the South-East Asia Region, World Health Organization.
Yosse, M.R., Nikzad, R., Nikzad, M., Mousapour, A., Ramazanpour, S.H., Rahimi, M.,
2014. High helmintic infection of the European grass snake, Natrix natrix and
the dice snake, Natrix tessellate (Serpentes: Colubridae) from Iran. Asian Pac. J.
Trop. Dis. 4 (Suppl. 1), S263S267.
Yousse, M.R., Hoseini, S.H., Mobedi, I., Ahoo, M.B., Rajabloo, M., 2010. Ophiotaenia
europaea, from Natrix natrix of north of Iran. World J. Zool. 5, 272274.
Yousse, M.R., Mousapour, A.A., Nikzad, R., Mobedi, I., Rahimi, M.T., 2013. First report
of Telorchis assula (Digenea: Telorchiidae) in three reptile species from North of
Iran. World J. Zool. 8 (3), 243244.
Zare Mirakabadi, A., Teymurzadeh, S., 2008. Venomous Snakes of Iran, Prevention,
First Aids and Treatment. Teymurzadeh and Tabib Press, Tehran.