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From Pastinachus Sephen (Myliobatiformes: Dasyatidae) from the Gulf of Oman Institute of Parasitology, Biology Centre CAS Folia Parasitologica 2017, 64: 014 doi: 10.14411/fp.2017.014 http://folia.paru.cas.cz Research Article Two new species of Tetragonocephalum (Cestoda: Lecanicephalidea) from Pastinachus sephen (Myliobatiformes: Dasyatidae) from the Gulf of Oman Atabak Roohi Aminjan and Masoumeh Malek School of Biology and Center of Excellence in Phylogeny of Living Organisms, College of Science, University of Tehran, Iran Abstract: In the present study two new species of Tetragonocephalum Shipley et Hornell, 1905, T. mackenziei sp. n. and T. kazemii sp. n., are described from the spiral intestine of the cowtail stingray, Pastinachus sephen (Forsskål), from the northern coast of the Gulf of Oman. Tetragonocephalum mackenziei is distinguished from the 16 other valid species of Tetragonocephalum by a unique combination of characteristics, i.e. sperm-filled seminal receptacle in immature proglottids, body length (7.7−17.5 mm), body width (213−288 µm), number of proglottids (34−49), number of testes (10−14), size of scolex (228−315 µm × 213−288 µm) and size of acetabula (56−73 µm × 61−75 µm). Tetragonocephalum kazemii is morphologically distinguishable from its valid congeners and T. mackenziei based on a combination of characteristics, including body length (28.8−36.6 mm), number of proglottids (50−65), number of testes (30−42), size of scolex (388−564 µm × 326−448 µm), size of acetabula (62−86 µm × 57−90 µm) and testes (25−39 × 21−32). This brings the total number of validly described species of Tetragonocephalum to 18 and expands our knowledge of this diverse genus to now include the Gulf of Oman, as well as Arafura Sea, northern Indian Ocean and western Pacific Ocean. Keywords: elasmobranchs, northern Indian Ocean, T. mackenziei sp. n., T. kazemii sp. n., sperm-filled seminal receptacle The genus Tetragonocephalum was erected by Shipley Malek (2016) described T. sabae Roohi Aminjan et Malek, and Hornell (1905) for the species T. trygonis Shipley et 2016 and T. salarii Roohi Aminjan et Malek, 2016 from Hornell, 1905 from Brevitrygon walga (Müller et Henle) Maculabatis randalli (Last, Manjaji-Matsumoto et Moore) from the Gulf of Mannar off the coast of Ceylon (now Sri from the Gulf of Oman. Lanka). Jensen (2005) revised the order Lecanicephalidea In addition to Tetragonocephalum, the order Lecanic- and evaluated the taxonomic status of Tetragonocephalum ephalidea currently includes 24 valid genera. The species and its members. In this monograph, 22 nominal species of Tetragonocephalum possess a dumb-bell-shaped uter- of Tetragonocephalum were examined, of which 14 were us in gravid proglottids, which appears to be unique to considered valid. In addition to these valid species, three this genus. Thus, this character could be considered as species were considered species inquirendae and five were the most useful discriminative feature to characterise the considered nomina nuda. Since 2006, five species have genus among other lecanicephalideans. There are some been described in the genus from Indian waters, namely, other diagnostic characteristics for Tetragonocephalum T. govindi Khamkar et Shinde, 2012; T. panjiensis Kham- such as acraspedote strobila, testes distributed anterior kar, 2011; T. pulensis Kankale, 2014; T. ratnagiriensis to the cirrus sac, ovary C-shaped in cross-section and Khamkar, 2012; T. sepheni Lanka, Hippargi et Patil, 2013 conspicuously enlarged genital atrium and pore (Ivanov (Khamkar 2011, 2012, Khamkar and Shinde 2012, Lanka et and Campbell 2000, Jensen 2005, Jensen et al. 2016). al. 2013, Kankale 2014). These five species are unavailable This study proposes two new species of Tetragonocepha- due to violation of Article 16.4 of the International Code of lum described from Pastinachus sephen (Forsskål) from Zoological Nomenclature (ICZN); according to this arti- the Gulf of Oman, Iran. These two new species clearly cle holotype, or syntypes, for the every new specific name possess the characteristics of the genus, although mor- published after 1999, must be deposited in a collection and phological differences exist between the two new pro- the location of that collection accompanied in the original posed species and all other validly described species of publication (ICZN 2016). Recently, Roohi Aminjan and Tetragonocephalum. Address for correspondence: M. Malek, University of Tehran, School of Biology and Centre of Excellence in Phylogeny of Living Organisms, College of Science, University of Tehran, Enghelab Ave., Tehran, 14155-6455 Iran. Phone: +98-21-61112702; Fax: +98-21-66405141; E-mail: [email protected] Zoobank number for article: urn:lsid:zoobank.org:pub:0B9639AF-491C-4BB1-ADC9-7D29DB2FFA7A This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. doi: 10.14411/fp.2017.014 Roohi Aminjan and Malek: Two new species of Tetragonocephalum MATERIALS AND METHODS ination and its voucher – partially measured). Worms A total of 15 specimens of Pastinachus sephen were collected 7.7−17.5 mm (11.8 mm; 3) long, apolytic; maximum width by the fisheries research vessel Ferdous in the northern waters 213−288 (253; 3) at scolex proper; 34−49 (40; 3) proglot- of Gulf of Oman in September 2010 (25°05'12''N; 60°02'16''E). tids (Fig. 1A). Scolex 228−315 (275; 3) long by 213−288 Each specimen was given a unique collection number (MM708, (253; 3) wide, consisting of scolex proper and apical organ. MM712, MM720, MM722, MM735−MM742, MM744, MM762 Scolex proper 124−178 (143; 3) long by 213−288 (253; and MM763). All host individuals were photographed and mor- 3) wide, bearing four acetabula. Acetabula sucker-like in phometric and morphological characteristics were recorded to form, appearing on surface of scolex proper at upper cor- inform species identification. Data and images of the host speci- ners, 56−73 (64; 3; 12) long by 61−75 (68; 3; 12) wide. mens are available at the elasmobranch host section of the Global Apical modification of scolex proper cylindrical, bearing Cestode Database at www.elasmobranchs.tapewormdb.uconn. apical organ. Apical organ muscular, 108−137 (124; 3) edu (Caira et al. 2012), accessible by the collection codes and long by 198−218 (205; 3) wide, non-invaginable, non-re- numbers. Species identity was confirmed using Naylor et al. tractable (Fig. 1B,C). (2012) and Henderson et al. (2015). Apical organ covered with tubercles suggesting glandu- Host individuals were dissected along the mid-ventral line; lar surface (Fig. 2B). Microtriches on apical modification spiral intestines were removed and opened by a longitudinal inci- of scolex proper and scolex proper (Fig. 2C) not observed. sion. Subsequently, spiral intestines were fixed in 10% seawater Strobila covered with capilliform filitriches (Fig. 2D). buffered formalin, shaken vigorously and held for approximately Cephalic peduncle absent. Proglottids acraspedote. Im- seven days. mature proglottids 25−36 (29; 3) in number, initially wid- Spiral intestines and intestinal contents were examined under er than long, gradually becoming longer than wide (3−16 a stereo microscope. Tapeworms were carefully removed from [8; 3] of immature proglottids longer than wide); most of the spiral intestine and washed in distilled water for about one them with seminal receptacle filled with sperm; two poste- hour before being preserved in 70% ethanol. Parasite specimens rior-most immature proglottids 239−391 (314; 3; 6) long were prepared as whole mounts for light microscopy observation by 163−192 (176; 3; 6) wide. Mature proglottids 2−3 (2; 3) according to Koch et al. (2012). in number; two posterior-most mature proglottids 337−516 Whole mounts were studied using a LEICA DM500 light (429; 3; 6) long by 153−222 (190; 3; 6) wide (Fig. 1D). microscope. Images of specimens of Tetragonocephalum were Gravid proglottids 7−10 (8; 3) in number; two posteri- taken using a LEICA ICC50 HD colour digital camera mounted or-most gravid proglottids 799−1,648 (1,161; 3; 6) long by on the LEICA DM500 light microscope (Buffalo Grove, Illinois, 170−230 (202; 3; 6) wide (Fig. 1E). USA) and measurements were taken using the image analysis Testes oval to spherical, 10−14 (12; 3; 6) in number, software Leica Application Suite (LAS EZ v.3.0.0). Measure- 17−32 (24; 3; 18) long by 18−38 (27; 3; 18) wide, restrict- ments were analysed in IBM® SPSS® Statistics Package v.22. All ed to anterior region of proglottid, extending from anteri- measurements of the reproductive organs were taken from ma- or margin of proglottid to anterior margin of anterolater- ture proglottids. Measurements are given in micrometres (µm) al fields of vitelline follicles, in two irregular columns in unless otherwise indicated and provided as the range followed dorsoventral view, arranged in multiple layers deep. Vas in parentheses by the mean, number of worms examined, and the deferens extending from level of anterior margin of ovary total number of measurements if more than one measurement was to cirrus sac, entering cirrus sac at distal end (Fig. 1D). Ex- taken per worm. Illustrations were prepared under the microscope ternal seminal vesicle absent. Internal seminal vesicle pres- with the aid of a drawing tube. ent, visible in gravid proglottids (Fig. 1E). Cirrus sac oval Some scoleces were prepared for ultrastructural studies us- in form, angled anteriorly, 56−77 (65; 3; 6) long by 47−90 ing scanning electron microscope (SEM) following the protocol (67; 3; 6) wide, containing coiled cirrus. Cirrus armed with of Jensen (2005). The specimens were sputter coated with ap- spinitriches. proximately 10 nm of gold/palladium and examined with a field Ovary oblong in dorsoventral view, incomplete ring- emission scanning election microscope (HIT4160102, Hitachi, shaped in cross-section, 91−145 (119; 3; 6) long by Tokyo, Japan) at the School of Electrical and Computer Engineer- 97−145 (125; 3; 6) wide, symmetrical.
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