New record of Thubunaea Pudica Seurat, 1914 (: Physalopteridae) in lizard mutabilis from Original Article Samar F Harras, Rasha A Elmahy Zoology Department, Faculty of Science, Tanta University, Tanta, Egypt

ABSTRACT Background: Studies on taxa remain poorly described in cold blooded , with rareness of data on the helminth community of Egyptian ones, especially lizards. The available literatures are mostly restricted to ecological studies rather than descriptive ones. Objective: To identify and give full description for that inhabit the Desert Agama, Trapelus mutabilis (T. mutabilis) caught from El-Dabaa desert, Egypt. Material and Methods: Nineteen Agama lizards having the characteristic morphological criteria of T. mutabilis

using light microscopy. Those subjected for scanning electron microscopy (SEM) were dried, coated and examined. Results:were dissected Seven andout examinedof nineteen for dissected parasitic lizardsinfection. were Gastrointestinal found to be infected nematodes with were the nematodecollected, fixed Thubunaea and identified pudica (T. pudica) (Family: Physalopteridae). They were collected from the stomach and small intestine of T. mutabilis. The main characteristics of adult T. pudica are: symmetrical anterior cephalic structure similar in both sexes, vulva

caudal papillae and two subequal stout spicules. Conclusion:is situated in the first tenth of the body, the tip of male tail ends beyondT. pudica well-developed using both light caudal and SEM.alae withMoreover, 32 true T. mutabilis lizard represents a new host record for T. pudica in a new geographic locality El-Dabaa desert as there are no reports of Thethis studyspecies added recorded the first from fully Egypt. described details for

Keywords: Egypt, El-Dabaa deserts, light microscope, , SEM, Thubunaea pudica, Trapelus mutabilis. Received: Accepted:

Corresponding13 June, Author: 2019, Samar F Harras,7 July, Tel. 2019. : E-mail: [email protected] Print ISSN: Online ISSN: 0020 Vol. 403271567, 12, No. 2, Ausgust, 2019. 1687-7942, 2090-2646, introduction Uromastyx Acanthocercus PseudoTrapelus Although more than 400 species of have Cuvier, 1817; Merrem, 1820; been reported in Egypt[1] there is minimal data on ofFitzinger, the 1843 Agama and, until Moody reviewedFitzinger, previously 1843. their helminth assemblage. Worldwide studies on proposedMany species generic of this names family for were species-groups. [7]classified as members the helminth communities of reptiles are limited to information regarding their ecology and The genus Trapelus rather than their detailed description . from northern Africa across the Middle East to Asia [2] consists of 15 species.Cuvier, Five species 1817 which occur is in distributed northern Lizards are good models for parasitological and Africa: T. mutabilis T. pallidus Reuss, ecological studies that describe their helminthic T. schmitzi infections, different feeding habits, activities and Merrem,T. tournevillei 1820 and Lataste, 1880 in . Previous studies 1833 in Egypt; T. savignii Wagner Dumeri and Böhme, and Bibron, 2007 demonstrated the high biodiverse[3] capacity of in and Niger; parasiticflexibility nematodes of habitat that location accompanies the varied thickAlgeria heads, and ; deeply andsunken tympana and a few spiny predispositions of lizards[4]. However, this parasite scales1837 in above Eastern the Egypt.ear opening They are. characterized by short fauna is rarely explored in lizards as a host, with respect [7] to the taxonomic aspects, compared to other groups of T. mutabilis lizard, is widespread, occurring across vertebrates[5,6]. northern Africa from the through , , Algeria, Tunisia, , and the to Egypt[8]. It differs from its close relative Agama Trapelus species, T. pallidus within the genus by possessing The lizard family Gray, 1827 is represented inPersonal Africa non-commercial by five genera: use only. PUJ Daudin, copyright 1802; © 2019. All rights reserved DOI: 10.21608/puj.2019.13643.1046

123 PARASITOLOGISTS UNITED JOURNAL irregular dorsal rhomboidal and scattered enlarged base of the tail. Coloration of the live specimen is sandy phosphate buffer saline and fixed overnight in 3% scales; but it has similar scales on the hind limbs and thenglutaraldehyde dehydrated in through 0.1 M phosphategraded series buffer of (pHethanol. 7.4) has violet blue throat . Dehydrationat 4°C. Specimens was performed were post in two fixed changes in 1% of absolute osmium gray with four to five [8,9]transverse bands. Male specimen ethanol and critical point dried. Specimens were The nematode family Physalopteridae constitutes mounted on stubs, coated with gold and examined with commonthree subfamilies: in reptiles, Thubunaeinae birds, mammals, Sobolev, rare in 1949, amphibians occurs a JEOL (5300 JSM) SEM at an accelerating voltage of 25 only in reptiles; Physalopterinae[10] Railliet, 1893, is K.V. African taxa of the genus Trapelus and absent in fish ; and ProleptinaeThubunaea Schulz, Seurat, 1927, of theIdentification collected nematode of the host, species followed[9] followed the key the forkey the of occurs mainlyPhysalopteroides in fish and rarely in reptiles. Two genera Anderson et al. and Gibbons[14]. ,All and measurements identification areparasites included of reptiles in Thubunaeinae; and are characterized by absence were given in millimetre.[13] of1914 a cephalic and ring, presence Wu of and many Liu, caudal1940. Theypapillae are with an ornamented cuticle forming papillary plates Ethical Consideration: Zoology Department’s council on male caudal surface[11]. The two genera differ from of the Faculty of Science, Tanta University, Egypt, each other by the symmetry of their cephalic structures approved the experimental design and the plan of work. which is symmetrical in Thubunaea, and asymmetrical in Physalopteroides[11]. results Pereira et al. pointed to the fact that the taxonomy of several nematodes[12] remain poorly described in T. mutabilis these cold-blooded animals in Egypt. Hence, full lizard examined for gastrointestinal parasitic infection, morphological studies of these parasites are important twenty-nineFrom seven nematode out of the specimensnineteen identified (eighteen females to complete the database and help clarify the current and eleven males) of T. pudica taxonomic confusion. Accordingly, the present study using light and SEM. Seurat, 1914 belonging to descriptive light and SEM picture of adult nematode familyFamily: Physalopteridae were identified and described specieswas planned affecting to give T. mutabilis for the first lizard time, caught a full from detailed EL- Subfamily: Thubunaeinae Dabaa desert, Egypt. T. pudica Physalopteridae Railliet, 1893

General description: Seurat, 1914 Adults (Figures are 1medium and 2). sized with material and methods

Nineteen Agama lizards caught from El-Dabaa Mouthfinely transverse is dorsoventrally striation elongated of cuticle and along surrounded their entire by twobody; symmetrical cephalic end lips. is rounded Each lip and is similarprovided in bothwith sexes.three rounded forwardly directed teeth on its inner side, with morphologicaldesert, Matrouh and (30°1′36′′N, characteristic 28°26′9′′E) picture of T. during mutabilis the a pair of submedian papillae and a small lateral amphid period(Reptilia: from Agamidae), September 2014were toexamined August 2015, for having parasitic the infection. The collected hosts were anesthetized using simple, and situated in the lateral cervical region slightly chloroform. The gastrointestinal tracts were removed between them (Figures 1B, and 2A). Deirids are small, leads into short vestibule (Figure 1A). Oesophagus is separately. Nematodes found in the stomach and composedbelow nerve of anterior ring level muscular (Figures part, 1A encircled and 2B). by Mouth nerve anteriorin 0.7% salineparts of solution small andintestine each were part wasselected. examined Live ring and posterior glandular part. Excretory pore opens specimens were washed in physiological saline and to the exterior, slightly anterior to the junction of both portions of oesophagus immediately behind the nerve Specimens were cleared in lactophenol then mounted infixed glycerin relaxed jelly in forwarm light 70% microscopical ethanol with examination 10% glycerol. and Malering (Figures 1A and 2C). identification.Specimens were measured and examined using (seven adult specimens): Body length: 5.25- 10.85; maximum width: 0.10-0.42. Vestibule length is microscope, and representative photomicrographs 0.03-0.07. Entire oesophagus measures 0.83-2.06 in Olympus CX31 microscope, and Zeiss Stemi 2000-C length, muscular portion is 0.1-0.2. Nerve ring is 0.03- were made with the aid of camera Lucida. 0.07, excretory pore is 0.2-0.22, and deirids are 0.16- were taken with E-330DC 7.4V digital camera. Drawings measuring0.17 from anterior 0.05, while end the of body,right onerespectively. measures There 0.04. Tipare For SEM, specimens were isolated carefully oftwo tail stout is pointed subequal distally spicules; with well-developed the left spicule caudal is longer alae and washed in isotonic saline solution, followed by

(0.02-0.03 wide) bearing numerous stout papilliform 124 Thubunaea Pudica in Trapelus mutabilis from Egypt Harras and Elmahy irregularly arranged cuticular ornamentation on one side and 5 on the other side. SEM revealed that (protuberances) on the ventral side. Caudal papillae papillae are rather similar in appearance, while light microscopy revealed them as pedunculated (Figures can be distinguished by their larger size and defined shape. There are about 32 papillae; 9 pairs precloacal 1C, and 2E). Minute lateral phasmidal pores are located and 14 postcloacal. Of the 14 postcloacal papillae 9 are slightly anterior to the tail projection (Figure 2F).

Fig 1. Thubunaea pudica. Camera Lucida. A, oesophagus, nerve ring (NR), deirid (D) and excretory pore (EP). B, anterior end of female, lateral view showing mouth (M), vestibule (V), muscular (MOe) and glandular (GOe) parts of (T), a pair of submedian papillae (SP) and a small lateral amphid (A). C, caudalenface endview of of male, cephalic subventral region viewshowing showing mouth cloaca (M) surrounded(C), caudal alae by two (CA), symmetrical caudal papillae lips; (CP),each pedunculatedlip has three rounded papillae, teeth (PP) protuberances (Pt) and two spicules (S). D, posterior end of female showing anal opening (AO) and small mucron (M). E, larvated egg, lateral view.

125 PARASITOLOGISTS UNITED JOURNAL

Female (seven gravid specimens): Length of body is is conical, rounded containing anal opening and

17.45-22.10 and maximum width is 0.7-1.05. Length Taxonomyterminating summary: in a small mucron (Figures 1D, and 2G). of vestibule is 0.04-0.042. Length of entire oesophagus Type of host: Desert Agama, T. mutabilis respectivelyis 2-3.35, of whichfrom theanterior muscular end portionof the bodymeasures (Figures 0.2- 0.25. Deirids and nerve ring are 0.2-0.21 and 0.16-0.3 Merrem, 1820. extremity as a transverse slit bound anteriorly and Collection site:time: El-Dabaa desert, Matrouh (30°1′36′′N, 1A, and 2B). Vulva is situated 2.05-3.7 from anterior Site28°26′9′′E). of infection: Stomach lumen and anterior part of is long and muscular. Two uteri contain numerous eggs small intestine. September 2014 to August 2015. withposteriorly larvae. by Eggs slightly are oval, swollen thick-walled, lips (Figure embryonated, 2D). Vagina Prevalence:

36.8% (7 out of 19 hosts examined), with and measure 0.03X0.04- 0.05X0.04 (Figure 1E). Tail mean intensity of 4.14 (1-17 worms/host).

Fig 2. Thubunaea pudica. SEM micrographs. A, amphid (A) on each lip. B and apical C, anteriorview of female,end of female, showing lateral two viewsymmetrical showing lips, deirids 3 rounded (D) and teethexcretory (T), apore pair (EP). of submedian papillae (SP) and small lateral D, E, caudal end of male, ventral view showing cloaca (C), caudal alae (CA), caudal papillae (CP) and protuberances (Pt). F, caudalanterior end end of ofmale, female, ventrolateral ventrolateral view view showing showing phasmid vulva (Ph). (V). G, posterior end of female, ventral view showing the anal opening (AO) and small mucron (M).

126 Thubunaea Pudica in Trapelus mutabilis from Egypt Harras and Elmahy

DISCUSSION biometric features, where the present female specimens

In our study, description of the discovered “Thubunaea” species is based on the generic characters 0.54are longermm). inThe length present (17.45-22.1 specimens mm also vs. have 15.64-19.17 longer given by Seurat[15] and Belle[16]. These characters are: oesophagusmm) and larger compared in width to that (0.7-1.05 described mm by vs. Moravec 0.381- mouth with two lateral simple lips, each lined with et al.[18] three small teeth in the internal surfaces and a pair of may be due to host variation and parasitological (2-3.35 mm vs. 1.93-2 mm). These differences relaxing agents), since all other general morphological malesubmedian caudal papillae; alae are no weIl lateral developed flanges; with cervical four papillaepairs of featurestechnique are (type the same. and concentration of fixative and immediately behind the nerve ring; short vestibule; pedunculated post-anal papillae and numerous sessile The number of male caudal papillae in the present pedunculated pre-anal papillae and four to five pairs of

papillae around the anus; two stout subequal spicules byspecimens Moravec is et 32 al which.[18] is in agreement with the type present; female posterior end terminating in a smallT. specieslarger specimen. of Seurat, This1914, difference but differs may from be that because reported the pudicaconical point; vulva is in the anterior fifth of the body; most anterior pairs whoof preanal detected papillae 35 papillae are usually in one two parallel uteri; oviparous eggs. Type species: poorly observable than others or may be hidden in The Seurat, morphological 1914. characterization and the wrinkles at this part. It is also concomitant with measurements of our nematode specimens are similar Sharpilo who reported that there is a probable to the original description of T. pudica reported from [19] several species of reptiles, including Scincus scincus, of caudal papillae in male Thubunaea members. The from North Africa[15]. In Egypt, T. pudica has been differenceintraspecific between variability SEM in the and number light andmicroscopy distribution of included in the checklist from the stomach of the snake sessile-pedunculated appearance in the caudal papillae Psammophis schokari by Myers et al. and from the observed in our study may be explained by their ability stomach of Scincus scincus in Abu Rawash[17] . for retraction. [17] In comparison with the previously reported Although more than twenty three species of T. pudica, our present specimen is identical to the Thubunaea are currently distributed all over the description by Belle[16] except for the length of the male world , their ultrastructural-topography has rarely been [20]studied using SEM, which provides detailed left spicule is 0.05 mm vs. 0.16 mm and right spicule is information to assess the validity of this genus. 0.04spicules, mm wherevs. 0.10 the mm. present male spicules are shorter; The difference in morphological and biometric measurements between the present T. pudica and the The present description somewhat resembles other reported species are illustrated in table (1). To the description of Moravec et al.[18], except for some the best of our knowledge, SEM studies are still scarce

Table 1. SEM characteristics of maleThubunaea species T. T. Thubunaea T. T. T. pudica ctenosaur mobedii spp. leonregagnonae eleodori (Present study) i[21] [22] [23] [24] [25] Cuticle Smooth With transverse Finely striation striation striation annulated transverse striation With fine cross With fine transverse With finely With a pair of With a pair With heavily With a pair With a pair of sub- Pseudo-labium With a pair of sub- sub-median of papillae on toothed outer of papillae median papillae median papillae papillae and outer margin margin and and 1 and 1 amphid and 1 amphid 1 amphid and 1 amphid amphid

Excretory 0.16 2 amphids pore site* Length 8.1 0.27 0.37 0.17 0.20 0.21 0.85 1.5 0.8 1.16 Oesophagus 13 6.19 10.25 6.9 No. of 1.29 16 Absent 10-16 16 pedunculated Data are not papillae available 12 for male No. of sessile 11-14 14 16 papillae Two17 equal 28 Two sub-equal Spicules Not spicules Absent Absent observed 0.066 and right 0.04 spicules; left 0.05 * From the anterior end, All measurements in mm

127 PARASITOLOGISTS UNITED JOURNAL 6. Poulin R, Leung TLF. Taxonomic resolution in T. ctenosauri from Ctenosaura pectinata in parasite community studies: are things getting forMexico membersT. ofmobedii this genus, from being Laudakia available nupta only in Iranfor five species;Thubunaea[21] sp. from Paraechinus aethiopicus in Central[22] Moody SM. Phylogenetic and historical ; T. leonregagnonae from Sceloporus; biogeographicalworse? Parasitol relationships2010; 137: 1967–1973. of the genera in the pyrocephalus [23]in Mexico and T. eleodori from 7. Family Agamidae (Reptilia Lacertilia). PhD Thesis. Liolaemus eleodori; in Argentina[24] . [25] 8. Baha El-Din S. A guide to the reptiles and amphibians ofUniversity Egypt. Cairo: of Michigan, The American Ann Arbor University 1980; 373 in pp.Cairo SEM morphological characterizations of both male andThe female present T. pudica study in providesEgypt. It shows for the the first accurate time symmetrical description of the anterior cephalic region, Press; 2006. as well as location of amphids, full description of caudal 9. AfricanWagner P,desert Melville lizards J, Wilms in TM,the SchmitzTrapelus A. mutabilis Opening region in males, including papillae distribution and a box of cryptic taxa: the first review of the North the position of phasmids in both sexes. Moreover, this T. pudica in T. mutibilis Merrem, 1820 complex (: Agamidae) lizard host in El-Dabaa desert as a new geographical 10. Andersonwith descriptions RC. Nematode of new taxa.parasites Zool Jof 2011; vertebrates. 163(3): localitystudy records in Egypt. for the first time 72- 88. nd Edition,

Acknowledgment: Deepest appreciations to Dr. Amal pp.Their development and transmission. 2 I. Khalil Professor of Parasitology for her kind revision 11. ChabaudCABI Publishing, AG. CIH Wallingford, keys to the Oxon,nematode UK 2000; parasites 650 of this work. Spirurida, part 1. Camallanoidea, Dracunculoidea, Author’s contribution: SF Harras shared in designing Gnathostomatoidea,of vertebrates. No. 3. Keys to generaPhysalopteroidea, of the order the study and research implementation, and in Rictularioidea and Thelaziodea. Commonwealth identifying the worms, reviewed the literatures, collected the data, and wrote the manuscript. RA Elmahy proposed the research idea, shared in collection descriptionAgricultural ofBureaux, the nematode Slough, Englandparasites 1975; of lizards: 27 pp. 12. StrongylurisPereira FB, oscari Luque JL, Tavares Luiz ER. Re- from Brazil and Pharyngodon mamillatus (Linstow, Conflictand identification of interest: of the worms. Travassos, 1923 (Heterakidae)

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