World Journal of Zoology 7 (1): 65-74, 2012 ISSN 1817-3098 © IDOSI Publications, 2012 DOI: 10.5829/idosi.wjz.2012.7.1.61177

Morphological and Molecular Studies of Sp. (: ) a Parasite of the Sandfish Lizard Scincus scincus Linnaeus, 1758 (Family: Scincidae)

Irene Sameh Gamil

Department of Zoology and Entomology, Faculty of Science, Helwan University, Cairo, Egypt

Abstract: Ophiotaenia sp. n. (: Proteocephalinae) is collected from the intestine of Scincus scincus Linnaeus, 1758 (Family: Scincidae) lizards from Sinai, Egypt. This is rare with both prevalence and intensity low. The medullary position of the vitellaria and reproductive organs, the uniloculate suckers and the two separate fields of testes placed it in the Ophiotaenia La Rue, 1911. It differs from all other Ophiotaenia species by the presence of excretory pores covering the whole worm and other special characters. Molecular data corroborated our findings at the morphological level. This is the first species of Ophiotaenia from Egyptian lizards and the first data on its phylogenetic position is provided.

Key words: Proteocephalidae Ophiotaeniidae Lizards Sinai Egypt

INTRODUCTION cyclophyllidean worms, mainly species of the genus Oochoristica [18-21], Nematotaenia mabuiae, Although numerous recent ecological studies are Cylindrotaenia allisonae and C. jaegerskioeldi [6] and available on helminth communities of , there is a Mesocestoides sp. tetrathyridia [18]. rareness of data on helminth communities of Egyptian The phylogeny of the order Proteocephalidea has ones especially lizards. Generally, reptiles harbor many been studied at generic and subfamiliar levels [22, 23] or cyclophyllidean tapeworms genera as Oochoristica Lühe, as a part of studies on the phylogeny of the in 1898 [1-4], Mesocestoides Vaillant, 1863 tetrathyridia [5], general, mainly in comparative morphology and Nematotaenia Lühe, 1910 and Cylindrotaenia Jewell, relationships among individual orders [24]. In addition to 1916 [6] in addition to proteocephalidean ones. morphological characters that are often variable, difficult Order Proteocephalidea Mola, 1928 is a very old to homologise or of little significance [25], molecular data group of helminths infecting mainly freshwater fishes, have been widely used in phylogenetic studies of and reptiles [7, 8]. The proteocephalidean tapeworms generally [26, 27] and proteocephalideans in genera recorded to infect reptiles were Proteocephalus particular [28-30] using several genes and developed Weinland, 1858, Crepidobothrium Monticelli, 1900, techniques as attempts in solving many taxonomic Acanthotaenia von Linstow, 1903 (Syn. Rostellotaenia problems. Freze, 1963), Ophiotaenia La Rue, 1911, Deblocktaenia Therefore, the aim of the present study was to Odening, 1963, Kapsulotaenia Freze, 1963, Tejidotaenia evaluate the phylogenetic position of the cestode Freze, 1965, Vaucheriella de Chambrier, 1987 and Ophiotaenia sp. n. parasite of the sandfish lizard Scincus Cairaella Coquille and de Chambrier, 2008 [7-13]. scincus Linnaeus, 1758 (Family Scincidae) inhabiting Sinai Regarding the lizards, they have been infected with in Egypt on the basis of morphological and molecular data many proteocephalidean species as Acanthotaenia among proteocephalideans. Up to date, no species of overstreeti [14], Tejidotaenia appendiculata [15], Ophiotaenia has been known to occur in reptiles from Kapsulotaenia sandgroundi [16], in addition to Cairaella Egypt; therefore Ophiotaenia sp. represents the first henrii and Ophiotaenia nicoleae [12], while from the species of Ophiotaenia from Egyptian lizards and we scincid lizards (Family Scincidae), only one species O. provide the first data on its phylogenetic position not yet greeri [17] was recorded up till now in addition to studied using molecular markers.

Corresponding Author: Irene Sameh Gamil, Department of Zoology and Entomology, Faculty of Science, Helwan University, Cairo, Egypt. 65 World J. Zool., 7 (1): 65-74, 2012

MATERIALS AND METHODS proteocephalidean species belonging to the genus Ophiotaenia. Sequence identity was checked using the A total of 160 sandfish scincid lizards S. scincus were Basic Local Alignment Search Tool (BLAST). collected from Sinai Governorate, Egypt during summer All regions of ambiguous alignment were removed 2010. Lizards were transported alive to the laboratory, from the dataset prior to phylogenetic analyses. they were euthanized, dissected and their helminths were Phylogenies based on the principle of maximum collected. Cestodes harboring the small intestine were parsimony were computed using Molecular Evolutionary collected, washed in physiological saline, fixed in 4 % hot Genetics Analysis software version 4.0 (MEGA4). 1000 formaldehyde solution, stored in 70% alcohol and bootstrap replicates were conducted to assess the examined with a light microscope. Scoleces and statistical support for the tree topology. proglottides were embedded in paraffin, longitudinal and transverse serial sections were cut at a thickness of 5-10 RESULTS µm. Whole mounts and serial sections were stained with haematoxylin and eosin and mounted in Canada balsam. Morphology: These small cestodes are slightly flattened For scanning electron microscope (SEM), some worms, dorsoventrally, oval in transverse sections and rounded one of them was cut at the level of gravid proglottides, at the posterior end. They are 12–15 mm (n = 10) long and were transferred to 4% gluteraldehyde, postfixed in 1% up to 0.08 mm wide. The strobila is acraspedote,

OsO4 , dehydrated through graded ethyl alcohol; critically anapolytic and consists of 14–20 proglottides (n = 8): 6–9 point dried, coated with gold and examined using a JEOL immature, 3–4 mature, 3–5 pregravid and 2 gravid (Fig. 1a). JSM–5200 SEM at an accelerating voltage of 25kV. The scolex is aspinose, very mobile and tetralobulate Microtriches were classified according to the terminology 41–50 mm wide (x = 46, n = 7, CV = 8.6%) and 25–32 proposed by Chervy [31]. All measurements are given in mm(x = 27.6, n = 7) long. It tapers anteriorly, with micrometres unless otherwise indicated. Abbreviations conspicuous wrinkles and folds so acquiring a rough used in the description are as follows: x = mean; n = appearance; neither a metascolex nor an apical organ is number of measurements; CV = coefficient of variability present but the scolex is covered in its apical part by an (SD/x x 100; in %). elevated mound of tegument (Fig. 1b) and provided with Cestodes intended for molecular analysis were fixed a concentration of cells of electron dense granular with 96% ethanol. Total DNA was extracted using the contents below apex, which may represent gland cells Analytik Jena Tissue kit for DNA isolation from tissue (Fig. 3a). The four suckers are spherical to oval in shape, samples. Primers used to amplify about 316 base pairs of uniloculate, 16–22 mm (x = 18.44, n = 9, CV = 11.85%) in the mitochondrial 16S rRNA gene were designed in the diameter and are laterally situated; the anterior end of laboratory of Biotechnology center, Faculty of Veterinary each sucker is drawn apically and shows a conspicuous medicine, Cairo University, on the basis of several transverse slit (Fig. 1b). The surfaces of the sucker, representative sequences for proteocephalidean species. scolex and strobila are all densely covered with filitriches The following primers were used: 16S R-378 (5'-ACY YAA (Fig. 1d), but no microtrichial polymorphism or modified GTC AAC ATC GAG GTA GC -3') and 16S F-53 (5'- AAT microtriches at any of the studied areas of this worm are RGC CGC AGT ATH RTG ACT GTG -3('. observed. Polymerase chain reaction (PCR) fragments were The immature and mature proglottides are longer than obtained using GenAmp® PCR System 9700 wide to wider than long (length/width ratio 1.24: 0.96), thermo-cycler with the following parameters: denaturation while, the pregravid and gravid ones are wider than long for 5 min at 95°C; denaturation 1 min at 94°C, annealing 1 (length/width ratio 0.74: 1.20) (Fig. 3c–f). Longitudinal min at 54°C, elongation 2 min at 72°C for 30 cycles and the inner musculature is well developed, often forming final elongation 10 min at 72°C. PCR products were irregularly-shaped bundles of separated muscles (Fig. 3f). checked for size on 0.8% agarose gel containing 0.5 µg of An identical arrangement of the osmoregulatory system ethidium bromide/ml. PCR fragments were directly is observed; two pairs of main collecting canals are sequenced by cycle sequencing using Macrogen ABI situated internal to the vitellaria. The ventral canals are 3730XL sequencer. The sequence data obtained in the slightly wider than the dorsal ones (Fig. 3c–f). A dense present study were deposited in the GenBank– EMBL net of narrow secondary canals is directed externally. under accession number JN671892. To determine the These canals open to the surface, through fine ducts, by position of this worm within the order Proteocephalidea, conspicuous pores covering all over the whole worm we aligned a part of the 16S rRNA gene sequence (316 bp) from the scolex to the last gravid proglottis long, with a representative of published (Figs. 1b–d, 2a, b, 3f).

66 World J. Zool., 7 (1): 65-74, 2012

Fig. 1: SEM of Ophiotaenia sp. n. a Whole worm. Bar 500 µm. b Enlarged part of the scolex. Bar 50 µm. c posterior proglottides. Bar 100 µm. d Filitriches covering the whole worm. Bar 5 µm. Fig. 3: Line drawings of Ophiotaenia sp. n. a Scolex with Abbreviations: ft filitriches, p pore, pz proliferation cells containing granular contents (cg) below apex. zone, s slit, sc scolex, st srobila, su sucker. Bar 10 µm b Egg Bar 10 µm c Dorsal view of an immature proglottis. Bar 50 µm. d Dorsal view of a mature proglottis. Bar 50 µm. e Dorsal view of pregravid proglottis. Bar 50 µm. f cross section at the middle region of a pregravid proglottis. Bar 10 µm. Abbreviations: ci cirrus, cs cirrus sac, do dorsal osmoregulatory canal, em embryophore, lm longitudinal musculature, mg Mehlis' gland, on onchosphere, ov ovary, p pore, te testis, ut uterus, vc vaginal canal, vd vas deferens, vi vitelline follicle, vo ventral osmoregulatory canal.

Testes are medullary, spherical to oval, 4–6 (x = 4.86, n = 8) in diameter, numbering about 20–40 (x = 28.73, n = 22, CV = 23.37%), in two lateral fields overlapped by osmoregulatory canals. They do not overlap cirrus-sac, vagina or vas deferens and usually reach vitelline follicles (Fig. 3c–e). Cirrus sac is pyriform, thin-walled 33–46 mm (x = 38.1, n = 10, CV = 9.86%) long by 15–20 mm (x = 16.86, Fig. 2: SEM of Ophiotaenia sp. n. a A part of pregravid n = 7, CV = 13.45%) wide, containing coiled ejaculatory proglottides. Bar 50 µm. b Enlarged pores. Bar 5 duct and invaginated cirrus. Cirrus sac represents 19–25% µm. c Enlarged genital atrium (ga) with papillae-like (x = 21.71%, n = 7, CV = 12.10%) of proglottis width. Vas outgrowths (ou). Bar 10 µm. d Egg. Bar 1 µm. deferens is thin-walled and coiled forming few loops Abbreviations: e egg, p pore. (Fig. 3c–e).

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Fig. 4: Nucleotide alignment of a 316 bp fragment from the mitochondrial 16S rRNA gene of the published Ophiotaenia species. Dots denote homology with O. gallardi.

Several papillae-like outgrowths are observed inside the genital atrium (Fig. 2c); genital pores irregularly alternating, are situated in the anterior quarter of proglottides. Ovary is medullary, bilobed with follicular lobes, 98–120 mm (x = 112.21, n = 14, CV = 6.94%) wide and occupied 63–73 % (x = 67.86%, n = 14, CV = 5.6%) of proglottis width. Mehlis's glands 7–9 mm (x = 8.13, n = 8, CV = 12.20%) in diameter, representing 9–11 % (x = 10.13%, n = 8, CV = 9.79%) of proglottis width (Fig. 3c–e). Vagina is anterior, or posterior to cirrus sac, thin walled. Vitelline follicles are medullary, arranged in two lateral fields near margins of proglottides and occupy almost 80–88% (x = 84.6, n = 18, CV = 3.4%) of the entire proglottis length, interrupted at level of cirrus-sac and Fig. 5: Phylogenetic tree of 16S rRNA sequence of vagina (Fig. 3c–e). Ophiotaenia species inferred by the maximum Primordium of uterine stem is medullary, visible in parsimony using bootstrapping and 1000 immature proglottides. Development of uterus replicates. demonstrates type 1 uterine formation, beginning with a

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Table 1: List of taxa included in the dataset with host, locality, maximum identity percentage with the present species and GenBank accession number Species Host Locality Max.% Iden. GenBank Accession Ophiotaenia sanbernardinensis Helicops leopardinus Paraguay 87 AJ389511 Ophiotaenia europaea Natrix maura Europe 86 AJ389520 Ophiotaenia gallardi Notechis scutatus Australia 86 AJ389516 Ophiotaenia grandis Agkistrodon piscivorus United States 85 AJ389514 Ophiotaenia ophiodex Causus maculatus Africa 85 AJ389493 Ophiotaenia jarara Bothrops jararaca Brazil 84 AJ389492 Ophiotaenia paraguayensis Hydrodynastes gigas Paraguay 84 AJ389506 Ophiotaenia sp. Scincus scincus Sinai, Egypt, Africa JN671892* * Sequence obtained in this study. straight uterine stem in immature proglottides occupying The present cestode worm Ophiotaenia sp. n. the most length of proglottis, till the formation of up to 9 infecting S. scincus lizards is characterized by the thin-walled ramified uterine branches on each side (n = 10) following features of taxonomic importance in the in pregravid and gravid proglottides (Fig. 3c–e). Eggs are classification of proteocephalidean tapeworms: spherical to oval, not contained in capsules (Fig. 2d). Ophiotaenia sp. is small in size (12–15 mm) being the Embryophore is bilayered; external layer is 9–11 in smallest one compared with almost all African species of diameter; the oncospheral membrane is 6–8 in diameter; Ophiotaenia that had registered a total length of the oncosphere is spherical up to 5 in diameter with three body between 50–600 mm (see de Chambrier et al. [13]). pairs of hooks (Fig. 3b). Freze [7] has cited that a large body size is rare within the Proteocephalidea and most of them attain lengths of up to Molecular Data: A comparison of the partial sequence of the 16S rRNA gene of the present tapeworm with those of 30–70 mm. other proteocephalidean cestodes, in a phylogenetic The scolex of Ophiotaenia sp. is very mobile, being context, provided further support for placing this species capable to be pushed out, forward or backward or laterally as a new one within Ophiotaenia thus confirming and retracted. This character is consistent to the primitive taxonomic conclusions based on morphological data. orders of Cestoda [32]. Neither an apical organ nor a In the phylogenetic tree obtained by maximum metascolex are recorded in the scolex of Ophiotaenia sp., parsimony analysis of the 16S rRNA sequence data set, a however, either muscular or glandular apical organ were close relationship between Ophiotaenia sp. and O. previously described in the anterior part of the scoleces sanbernardinensis parasite of South American snake of O. nicoleae [12] and O. gallardi [33] geckos and snake Helicops leopardinus (Colubridae) is clear (Fig. 5) with a parasites, respectively. On the other hand, absence of maximum identity of 87% (Table 1) but with 30 different apical organ in Ophiotaenia sp. is in agreement with most base pairs with a percentage divergence of 9.5% (Fig. 4). African species of Ophiotaenia [13]. Ophiotaenia. sp. is with both prevalence and Reptilian proteocephalidean genera exhibit a intensity values low; being 12 % and 1-2/ host, gradation, from the simple uniloculate suckers in respectively. Ophiotaenia, which is the case in Ophiotaenia sp., notched uniloculate suckers in Crepidobothrium, DISCUSSION biloculate suckers in Tejidotaenia to tetraloculatate Tapeworms of the order Proteocephalidea Mola, 1928 suckers in Deblocktaenia [32]. are diagnosed by the following features: strobila small to The transverse slit in the anterior margin of the medium in size, distinct external segmentation, suckers of Ophiotaenia sp. was never recorded parenchyma divided into cortical and medullary regions previously in any proteocephalidean species and based generally separated by internal longitudinal musculature, on their localization; they may be openings of gland cells one set of gonads located medullary or in combination in with secretions participating in the attachment of the medulla and cortex per segment, numerous testes scolex between the villi of the host intestine. Bru anská (between 20-400), a lateral genital pore, a bilobate ovary at et al. [34] have recorded previously, using transmission the posterior end of proglottis, follicular and lateral electron microscopy (TEM), secretory products at the vitellaria and a variable scolex with four suckers, in sucker edges of the proteocephalidean P. longicollis addition, an apical organ, such as a sucker, or rarely an coming from scolex glands but with no special pores in armed rostellum may be present [8, 28]. the sucker.

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The scolex of Ophiotaenia sp. is with a concentration the fine pores might be excretory ones. Scattered of cells with granular contents below the apex, which may excretory openings were also observed in the posterior represent gland cells explaining the observations of bladder surface of the cyclophyllidean Taenia crassiceps sucker's pores. Similar granular contents were recorded in cysticercus [49] with a suggestion that they may help in O. bonneti [35] as well as many species of osmoregulation. Proteocephalus [36, 37]. Befus and Freeman [38] have Proteocephalidean parasites of reptiles have a explained earlier that such cells with granular contents relatively stable, species-specific number of testes [13]. may play a role with their secretions during the The testes of Ophiotaenia sp. are present in two field attachment of the worms to the host intestine. arrangements. Similar observations were recorded in the Furthermore, evident scolex glands with ducts snake parasites O. georgievi [13] and O. gallardi [33] and transporting their secretory products to the tegument in addition to C. henrii [12] and K. sandgroundi [16] lizard surface were recorded in the proteocephalideans parasites. Freze [7] has previously differentiated the P. ambloplitis [39], P. joanae [40], P. percae, P. torulosus genus Ophiotaenia from Proteocephalus essentially by and P. exiguus [41] and Silurotaenia siluri [42] with the the presence of two testicular fields and preformed uterus. same conclusion of attachment function of the worm to The position of the ovary of Ophiotaenia sp. is basal the host tissues. Rego [43] have concluded that the apical which is commonly found in Proteocephalidea [37, 50], glands are more frequent in proteocephalidean species of with few exception [15] and the uterus is medullary, Ophiotaenia, Proteocephalus and Nomimoscolex; which although a cortical position of uterine stem was also are all genera without metascolex. recorded [10]. Hanzelová and Špakulová [25] have The amplification of the surface of Ophiotaenia sp. explained that the number of uterine branches in a gravid is achieved by the presence of filitriches which, according segment is a characteristic item and almost used as a to d’árska et al. [44] and d’árská and Nebesá ová [45] principle taxonomic criterion in proteocephalideans. As is magnify the absorption area and thus facilitate uptake of generally known, many proteocephalidean species have nutrients and agitate the microenvironment adjacent to between 5 and 12 uterine branches [51], although this the tapeworm to enhance nutrients and waste products in number varied considerably within the Proteocephalidea a state of flux across the tegument surface. [37, 50]. Generally, osmoregulatory canals in cestodes consist Within the Proteocephalidea, the uterus, in its of a pair of ventral and a pair of dorsal canals situated in immature and mature stages, shows two distinct and the medullary parenchyma [46]. In the Proteocephalidea, fundamentally different types of development (type 1 and the ventral osmoregulatory canals are always wider than type 2) [30], this character may become important in the dorsal ones [33-36]. From the ventral canals, proteocephalidean systematics, as previously suggested secondary osmoregulatory ones extend mainly in [10]. In the present study, type 1 uterine formation is proglottides and rarely in the scolex and neck parts demonstrated in Ophiotaenia sp. sharing many species of [40, 41, 47]. These canals then either terminate beneath the Ophiotaenia [12, 13, 33]. tegument [16, 42, 47] or open outside [36, 37]. In the In the present study, small papillae-like projections present study, similar observations are recorded with are recorded in the genital atrium of Ophiotaenia sp. Such terminal osmoregulatory canals opening outside by fine projections were neither recorded in any pores scattered all over the whole worm and not proteocephalidean nor any cestode. Medullary vitelline specified to only the scolex or the strobila. Depending on follicles of Ophiotaenia sp. agree with those recorded in their location, these pores may be of excretory functions. most species of Ophiotaenia [12, 13, 33, 35]. The presence of such pores covering the entire worm in The construction of the eggs of Ophiotaenia sp. is in the present study represents a unique feature for the agreement with those of all African taxa described until Proteocephalidea and is also rare within the Cestoda. now possessing two layered embryophore [7], but are not Similar fine pores, besides a large central one, were contained in clusters. de Chambrier [16] has recorded the recorded only within the scolex' ridges of the presence of egg clusters in K. sandgroundi parasite of proteocephalidean Corallobothrium solidum infecting Varanus komodoensis lizards and added that the majority the electric [48] but all of Australian proteocephalideans from varanids and described as openings of frontal glands participating in snakes possess eggs in clusters, although, Rego [32] has attachment and nutrition; depending on their sizes, this previously informed that in Proteocephalidea, eggs are large central opening can be that of frontal gland, while usually singly and egg capsules are rare.

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From morphological point of view, the new species of Proteocephalinae) but differ from all other the present study is positioned in the genus Ophiotaenia proteocephalidean cestodes hitherto described by several La Rue, 1911 (subfamily Proteocephalinae), depending on particular characteristics: the medullary distribution of gonads and vitellaria, the unarmed scolex, the uniloculate suckers as well as the two The presence of excretory pores covering all over the separate fields of the testes [7, 9]. whole worm. Ninety-six species of Ophiotaenia parasitizing The transverse slit in the anterior margin of the reptiles and amphibians are currently recognized as valid suckers. [9, 12, 13, 33, 52]; out of these, about 15 species are found The small papillae-like outgrowths in the genital in Africa, no one of them being from a lizard. World wide, atrium. only four species of Ophiotaenia were recorded from the lizards; these are the Australians O. striata Johnston, ACKNOWLEDGMENTS 1914 and O. amphiboluri Nybelin, 1917 infecting Lialis burtonii and Pogona barbata, respectively, in addition to I'm indebted to Dr. Hussein Ali Chairman of Virology O. greeri [17] from Sphenomorphus aignanus in Papua Department, Faculty of Veterinary Medicine, Cairo New Guinea and O. nicoleae Coquille and de Chambrier, University for his valuable advice and help with the 2008 from the gekkonid lizard Thecadactylus rapicauda phylogenetic analyses and to all members of the in Ecuador [12]. Biotechnology center. The results of the study of de Chambrier et al. [30] have previously confirmed the basal position of the REFERENCES parasites of monitors (lizards) as Acanthotaenia, Rostellotaenia and Kapsulotaenia considering them as 1. Conn, D.B., 1985. Life cycle and postembryonic the most basal taxa in the phylogenetic tree and they development of Oochoristica (Cyclophyllidea: concluded that type 1 uterine development appears to be Linstowiidae). J. Parasitol., 71: 10-16. primitive as it is present in the Acanthotaeniinae, the 2. Okafor, F.C., 1988. Oochoristica agamae Baylis, 1919 Gangesiinae and proteocephalideans from viperids and (Eucestoda, Linstowiidae) in one and two bat elapids. As a conclusion, we can say that Ophiotaenia species from Nsukka (Anambra State, Nigeria). Misc. sp. may represent one of the primitive species of Zool., 12: 11-15. Ophiotaenia depending on morphological characters. 3. Criscione, C.D. and W.F. Font, 2001. Development Brooks [22] and de Chambrier [10] have considered and specificity of Oochoristica javaensis (Eucestoda: Ophiotaenia and Proteocephalus synonymy and in Cyclophyllidea: Anoplocephalidae: Linstowiinae). phylogenetic analyses, Zehnder and Mariaux [28] and Comparative Parasitol., 68: 149-155. Hypša et al. [24] demonstrated their polyphyly. It has 4. Mašová, S., F. Tenora, V. Baruš and P. Koubek, 2010. been concluded that almost all proteocephalidean A new Anoplocephalid (Cestoda) from Tarentola cestodes found in reptiles and amphibians are members of parvicarinata (Lacertilia: Gekkonidae) in Senegal Ophiotaenia [7, 12, 35, 52]. de Chambrier et al. [35] and (West Africa). J. Parasitol., 96: 977-981. Ammann and de Chambrier [52] added that most species 5. McAllister, C.T., J.E. Cordes, D.B. Conn, J. Singleton of Ophiotaenia are strictly host specific infecting only and J.M. Walker, 1991. Helminth parasites of one species of definitive host. unisexual and bisexual Whiptail lizards (Teiidae) in de Chambrier et al. [30] suggested, as was also North America. V. Mesocestoides sp. Tetrathyridia proposed by Brooks [22], Rego et al. [23] and Zehnder (Cestoidea: Cyclophyllidea) from four spcies of and Mariaux [28], that colonisation of terrerstrial hosts Cnemidophorus. J. Wildlife Diseases, 27: 494-497. (reptiles and mammals) occurred on several occasions 6. Jones, M.K., 1987. A taxonomic revision of the during the history of Proteocephalidea. Nematotaeniidae Lühe, 1910 (Cestoda: We concluded that the morphology of Cyclophyllidea). Systematic Parasitol., 10: 165-245. Ophiotaenia sp. studied by light microscopy, SEM and 7. Freze, V.I., 1965. Essentials of Cestodology. Vol. V. histological observations as well as the sequence of its Proteocephalata in Fish, Amphibians and Reptiles. 16S rRNA gene obtained in this study, clearly Nauka, Moscow, 538 pp (In Russian; English demonstrate that this species should be considered to be translation, Israel Program of Scientific Translation, a member of Ophiotaenia (Proteocephalidae: 1969, Jerusalem, pp: 597).

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