Descriptive Osteology of Barbus Cyri (Teleostei: Cyprinidae) from Southern Caspian Sea Basin

Descriptive osteology of Barbus cyri (Teleostei: Cyprinidae) from southern Caspian Sea basin

Pariya JALILI1, Soheil EAGDERI1*, Nasrin NIKMEHR1, Yazdan KEIVANY2

1Department of Fisheries, Faculty of Natural Resources, University of Tehran, Karaj, Iran. 2Department of Natural Resources (Fisheries Division), Isfahan University of Technology, Isfahan, 84156-83111, Iran. Email: [email protected]

Abstract: The present study was conducted to provide a detailed descriptive osteology of Barbus cyri from Southern Caspian Sea for the first time and comparing it with those of B. lacerta from the Tigris and Urmia lake basins. For this purpose, twelve specimens of B. cyri were collected from Safidrud River using electrofishing device and fixed in 10% buffered formalin. Then, the specimens were cleared and stained with alcian blue and alizarin red for osteological examination. A detailed description of the osteological features of B. cyri was provided. Based on the results, several differences were observed between the two species. Barbus cyri can be distinguished from B. lacerta by having three pharyngobranchial, a consumptive neural spine of second centrum and having 3rd and 4th vertebra with posterior position of neural spines. Keywords: Barbus, Kura barbel, Cyprinidae, Caspian Sea, Skeleton, Bone.

Introduction has a wide distribution in the southern Caspian Sea Over the years, the generic name Barbus (Cuvier and basin (Berg 1949; Bianco & Banarescu 1982; Kiabi Cloquet 1816) has been used for about 800 nominal et al. 1999; Abdoli 2000; Esmaeili et al. 2014a, b; species of cyprinid fishes in Europe, Africa and Asia. Motamedi et al. 2014). Berg (1948-1949) refers As presently understood, however, it is split into Caspian Sea basin specimens to Barbus lacerta cyri. several distinct genera (Coad 2015). This genus is But, this subspecies recognized as a full species by characterized by a rounded or compressed body of Naseka & Bogutskaya (2009). moderate to very large size, large to very small scales Until now, the phylogenetic relationships of the (no scale sheath around the anal fin), the presence of members of this group is based on external barbels in most species, usually two pairs, often one morphological characters (i.e. corporal proportions, pair and sometimes none (and individually variable scale and ray number), and few interna1 ones (i.e. within species), the last unbranched ray in the short number of pharyngeal teeth and gill rakers) (Doadrio dorsal fin (usually 7-8 branched rays but sometimes 1990). Since, osteological characters can provide more) is thickened and spine-like and may bear teeth valuable information in taxonomy and phylogenetic or be smooth and a short anal fin, usually with 5 relationships in fishes (Keivany & Nelson 1998, branched rays (but some have 6) (Coad 2015). 2004, 2006; Diogo & Bills 2006; Keivany 2014a, b, There are 16 formerly recognized species of the c, d), therefore, the present study was conducted to genus Barbus in Iranian inland waters (Coad 2015); provide a detailed descriptive osteology of B. cyri only two species including B. cyri (De Filippi 1865) from southern Caspian Sea for the first time and and B. lacerta (Heckel, 1843) is now assigned to this comparing it with those of B. lacerta from Tigris and genus. Barbus lacerta is found in the Tigris and Urmia lake basins. The results will provide a basis for Urmia Lake basins of Iran (Coad 2015). Barbus cyri further phylogenetic study of Iranian members of the 105

Iranian Journal of Ichthyology (June 2015), 2(2): 105-112

Fig.1. Lateral view of Barbus cyri from Safidrud River.

Barbus group using osteological data. part of the vomer and the anterior part of the parasphenoid. The posterior part of the vomer is Materials and Methods pointed and its anterior part has a V-shaped notch. Twelve specimens of B. cyri were collected from The anterior part of the vomer is thicker than its Safidrud River (10.74±2.64mm; SL±SD) using posterior part. Two small semicircular preethmoid-I electrofishing device and fixed in 10% buffered are present in the antero-lateral part of the vomer formalin, after anesthetizing in 1% clove oil (Fig. 1). (Fig. 2c). The lateral ethmoid is located as a wall The specimens were cleared and stained with alizarin between the ethmoid and orbital regions; this bone is red S and alcian blue according to the protocol of concaved dorsally and flattened ventrally and its Taylor & van Dyke (1985) for osteological ventral surface bears many small pores. A cylindrical examination. The cleared and stained specimens kinethmoid exists between the maxillary bones (Fig. were studied using a stereomicroscope (Leica MC5); 3a). and their skeletal elements were dissected and The orbital region comprises the frontals, scanned by a scanner equipped with a glycerol bath parasphenoid, ptersphenoids, orbitosphenoids and (Epson V600). Drawing of the specimens were circumorbital series. The frontal is a large bony performed using CorelDraw X6 software. The element of the skull roof with a serrated anterior terminology of skeletal elements follows Rojo (2010) edge; it bears a mid-lateral pointed process. The and Howes (1982). The detailed osteological features supraorbital canal is enclosed by the lateral margin of of B. lacerta from Tigris and Urmia lake basins were the frontal. The two orbitosphenoids are fused via provided by Jalili et al. (2015) and Razavipour their ventral process that is also connected to the (2013). parasphenoid (Fig. 2b). The ptersphenoid is concaved in shape with two ventral processes for Results connecting to the parasphenoid. The ptersphenoid is The ethmoid region of the neurocranium consists of dorsally attached to the frontal, anteriorly to the the preethmoid-I, lateral ethmoid, supraethmoid, orbitosphenoid and posteriorly to the sphenotic. The vomer, kinethmoid and nasal bones (Fig. 2a, b). The anterior half of the parasphenoid is type-shape with a supraethmoid consists of two vertical and horizontal serrated anterior rim and its posterior half has a sections; the horizontal section bears two small triangular structure that its posterior margin anterior processes and a shallow middle depression bifurcates via a deep groove (Fig. 2c). with a cartilaginous lateral margins (Fig. 2a). The In the circumorbital series, the number of the vertical section of this bone is situated on the dorsal infraorbital bones was different in studied specimens.

106 Jalili et al.-Descriptive osteology of Barbus cyri

of Cobitis avicennae

Fig.2. Dorsal (a), lateral (b) and ventral (c) views of the neurocranium in Barbus cyri. Abbreviations: Bo: basioccipital; Epo: epiotic; Exo: exoccipital; Fr: frontal; Fon: fontanel; Le: lateral ethmoid; Nas: nasal; Orb: orbitosphenoid; Pa: parietal; Pe: preethmoid I; Ptt: posttemporal; Pr-Pp: posterior pharyngeal process; Pro: prootic; Ps: parasphenoid; Pts: pterosphenoid; Pto: pterotic; Stt: supratemporal; Scl: supracleithrum; So-cr; supraoccipital crest; So: supraorbital; Soc: supraoccipital; Sp: sphenotic; Se: supraethmoid; Vo: vomer. There are 4-6 infra-orbital and one supra-orbital the frontal. The suborbital canal is enclosed by the elements. The first circumorbital i.e. lacryamal, is the infraorbital bony elements (Fig. 3c). largest element of this series. The supraorbital is The otic portion of the neurocranium includes oval-shaped and located at the antero-lateral part of five bones viz. the parietal, epiotic, sphenotic, 107 Iranian Journal of Ichthyology (June 2015), 2(2): 105-112

Fig.3. Internal view of the upper (a) and lower jaw (b) and circumorbital series (c) of Barbus cyri. Abbreviations: An: Angular; Crb: coronoid process; Cm: coronomeckelian; Dn: dentary; Io 2-6: infraorbitals 2-6; Keth: kinethmoid; Ra: retroarticular; Mx: maxillary; Pmx: premaxillary; Mdip: maxillary descending process; Mdp: maxillary distal process; Mdcb: maxillary dorsal concaved border; Mdip: maxillary mid-lateral ascending process; Rap: rostral ascending process; Top: tail of premaxillary. pterotic and prootic. The postero-lateral margin of The upper jaw is made up of the maxillary and the parietal covers the dorsal part of the pterotic and premaxillary. The two maxillaries are located at the epiotic. The epiotic is oval-shaped with a posterior dorsal side of the premaxillary; this bone possesses a process (Fig. 2a); it is situated between the pterotic mid-lateral ascending and an anterior descending and supraoccipital. The pterotic has a trapezoid shape processes; it has also a distal process and a fossa on (Fig. 2b) and its postero-lateral part is well- its dorsal border. The premaxillary is wide and its developed. The sphenotic possesses a lateral process anterior part has a rostral ascending process (Fig. 3a). which connects the middle process of the frontal (Fig. The lower jaw is composed of four bones 2a). The ventral margin of the sphenotic is connected including the dentary, angular, retroarticular and to the prootic. The two prootics are connected to each coronomeckelian. The coronoid process, situated on other ventrally and to the parasphenoid dorsally by a the dorso-middle part of the dentary, is oriented descending process. This bone has two pores on its posteriorly. The middle part of the angular is wide anterior part. and its posterior part bears an articulatory facet. The The occipital region is composed of the retroarticular is a small bone situating under the supraoccipital, exoccipitals and basioccipital. The posterior part of the angular; the posterior part of the supraoccipital is pentagon in shape and has a blade- retroarticular is connected to the interopercle via the shaped crest. The exoccipital bears a large foramen retroarticular-interopercle ligament. A small and on its middle part. In the dorsal part of the elongated coronomeckelian is observed on the basioccipital, there are a pointed pharyngeal process medial face of the angular (Fig. 3b). and a concaved masticatory plate pointing laterally The suspensorium is formed by the (Fig. 2c). hyomandibular, ectopterygoid, endopterygoid, 108 Jalili et al.-Descriptive osteology of Barbus cyri

of Cobitis avicennae shape with a dental formula of 2.3.5-5.3.2 (Fig. 5a). The hyoid arch consists of the paired epihyals, hypohyals and ceratohyals and the unpaired urohyal and basihyal, and three pairs of the branchiostegal rays. The urohyal has two vertical and horizontal sections. The posterior margin of its horizontal section is pointed and its middle part is wider. In addition, the anterior part of its horizontal section is bifurcated. A small interhyal locates at the dorsal part Fig.4. The suspensorium and opercular series (Right side) (c) in Barbus cyri. Abbreviations: Ect: of the epihyal. The posterior part of the ceratohyal is ectopterygoid; End: endopterygoid; Hy: wider than its anterior part; its anterior part is hyomandibulare; Iop: interopercle; Mtp: bifurcated and attached to the dorsal and ventral metapterygoid; Op: opercle; Opp: opercular prominent process; Opj: opercular joint; P: palatine; Pop: hypohyals. The basihyal is T-shaped situating preopercle; Q: quadrate; Sj: spine and socket joint; between the hypoyals (Fig. 5b). Sop: subopercle; Sym: symplectic. The pectoral girdle consists of the cleithrum, metapterygoid, symplectic, quadrate and palatine supracleithrum, coracoid, mesocoracoid, scapula, (Fig. 4). The hyomadibular bone is almost triangular posttemporal, supratemporal and radials (Fig. 5a). in shape and its posterior part bears two The cleithrum bears two wide descending and protuberances; the ventral part of the hyomandibular ascending portions; its ventral part bears an antero- is connected to the interhyal. The ectopterygoid, medial downward process connecting to the anterior endopterygoid, metapterygoid, symplectic and part of the coracoid. Also, it has a dorsal process in quadrate form a bony complex connecting to the its ascending part for positioning the supracleithrum. anterior part of the neurocranium via the palatine. A The posterior part of the coracoid is wider than its long symplectic is enclosed by the metapterygoid anterior part and bears an ascending process for posteriorly, by the quadrate anteriorly and by the connecting to the mesocoracoid. The medial part of preopercle posteriorly. The anterior part of the the coracoid is bended ventrally and has a small pore. palatine bears three processes and a slightly deep A semicircular scapula is located between cleithrum depression for connecting to the vomer and and coracoid bones; this bone bears a large foramen preethmoid-I. and articulated to the first unbranched ray. The The opercular series is consists of four bones. ventral part of the mesocoracoid is V-shaped and The antero-dorsal part of the opercle has an opercular attached to the coracoid and scapula; the dorsal part process and a socket joint that is connected to the of the mesocoracoid is broadened and attached to the hyomandibular. A L-shaped preopercle presents in medial face of the cleithrum. The pectoral fin bears the anterior side of the opercle; its vertical part is four radials; the lateral radial is the thickest and three longer than that of its horizontal part. The ventral part others are long and flat. of the preopercle overlaps the dorsal part of the Pelvic girdle includes the paired pelvic bone, interopercle. The subopercle has an antero-dorsal pelvic splint and radials (Fig. 5b). The anterior part process pointed upwardly (Fig. 4). of the pelvic bone is U-shaped; this bone has a The branchial apparatus includes five type bones posterior long and a mid-lateral processes. The two including five pairs of the ceratobranchials, four pairs L-shaped pelvic splint is located at the postero-lateral of the epibranchials, three pairs of the hypobranchials side of the pelvic bone. There are three radial bones and the pharyngobranchial and three unpaired in the pelvic girdle. basibranchial. The fifth ceratobranchial is crescent in The Weberian apparatus is formed by four 109 Iranian Journal of Ichthyology (June 2015), 2(2): 105-112

Fig.5. Medial view of pectoral girdle (left side) (a) and pelvic girdle (b) of Barbus cyri. Abbreviations: Cl: cleithrum; Co; coracoid; Mco: mesocoracoid; Mlp; mid-lateral process; Pb: pelvic bone; Pcl; postcleithrum; Pop: posterior process; Ps: pelvic splint; R: radials; Sc: scapula.

Fig.6. Lateral view of the Weberian apparatus (a), dorsal anal fins (b) and caudal skeleton in Barbus cyri. Abbreviations: C15-28: centrum 15-18; Dfs: dorsal fin spine; Dpt: distal pterygiophore; Epu: epural; Hp 1-6: hypural plates 1-6; Hsp: hemal spine; Mtp: medial pterygiophore; Na 4: neural arch 4; Ns: neural spine; Pr 4: pleural rib 4; Ppt: proximal pterygiophore; Ph: parhypurale; Pls: pleurostyle; Rna: rudimentary neural arch; Sn: supraneural; Sty: stay. anterior centra with their ossicles including the oriented posteriorly. The two ventral processes of the tripus, intercalarium, scaphium and claustrum (Fig. fourth vertebra are fused to each other; the neural 6a). The tripus is positioned at the latero-ventral side arch of this bone is short and connected to the neural of the second vertebra and its dorsal part extends to arch and the supraneural of the third centrum. There the posterior part of the pleural rib of the fourth are 42-44 vertebra including 19-22 cranial and 20-24 centrum. The intercalarium is T-shaped and attached caudal centra. to the tripus and scaphium via the scaphia-intercalar The dorsal fin bears four unbranched and eight and intercalo-tripodal ligaments. The clastrum is branched rays, nine pterygiophores and one stay (Fig. positioned on the dorsal part of the scaphium; they 6b). The dorsal margin of 4th unbranched ray is meet each other via a clastro-scaphial ligament. The dentated. Two first pterygiophores are the largest and ventral portion of the pleural rib of 2nd centrum supports unbranched rays. The first pterygiophore is 110 Jalili et al.-Descriptive osteology of Barbus cyri

of Cobitis avicennae next to the 15th or 16th vertebra. In front of the dorsal is formed by five vertebra. In addition, in B. lacerta fin 9-10 free supraneural bones are observed. A of the Urmia lake basin, one stay is present in anal fin triangular stay bone supports the last branched ray. versus two stays of B. cyri and B. lacerta of the Tigris The anal fine originates at 28th centrum. This fin basin (Razavipour 2013). has three unbranched and five branched rays and is supported by six pterygiophores and two small stay Acknowledgments bones (Fig. 6b). We would like to thank M. Zamani Faradonbeh and The caudal skeleton consists of four centra with M. Nasri for their help during the fish collection. This the epural, parhypural, pleurostyle, uroneural and six study was financially supported by the University of hypurals bones (Fig. 6c). The pleurstyle is fused to Tehran. the last centrum. The neural arch of the second centrum is consumptive in some specimens. The first References hyporal is the largest one and attached to the Abdoli, A. 2000. The Inland Water Fishes of Iran. Iranian parhyporal. The epural is a long bone positioning at Museum of Nature and Wildlife. Tehran, 378 pp. the dorsal part of the neural arch of the first vertebra. Berg, L.S. 1948–1949. Freshwater fishes of the USSR and adjacent countries. Israel Program for Scientific Discussion Translations, Jerusalem. Berg, L.S. 1949. Freshwater fishes of Iran and adjacent There are little differences in the caudal and anal countries. Trudy Zoologicheskogo Instituta structures, Weberian apparatus, branchial arch and Akademii Nauk SSSR 8: 783-858. the numbers of the vertebra and supraneural elements Bianco, P.G. & Banarescu, P. 1982. A contribution to the between Iranian members of the genus Barbus. knowledge of the Cyprinidae of Iran (Pisces, Barbus cyri bears three pharyngobranchials, whereas Cypriniformes). Cybium 6: 75-96. B. lacerta bears two pharyngobranchial (Razavipour Coad, B. 2015. Fresh water fishes of Iran. 2013). The number of the supraneural is variable in www.briancoad.com. Retrieved 9/3/2015. the specimens of this genus; in specimens of Doadrio, I. 1990. Phylogenetic relationships and B. lacerta from the Tigris and Urmia lake basins, classification of western palaearctic species of the there are 7-8 and 7-10 supraneurals (Razavipour genus Barbus (Osteichthyes, Cyprinidae). Aquatic 2013), respectively, whereas B. cyri has 9 Living Resources 3(4): 265-282. Diogo, R. & Bills, R. 2006. Osteology and myology of the supraneurals. cephalic region and pectoral girdle of the South Barbus cyri bears 42 vertebrae and its fourth African Catfish Austroglanis gilli, with comments vertebra has neural prezygapophyses and neural on the Autapomorphies and phylogenetic postzygapophyses, whereas in two other population relationships of the Austroglanididae (Teleostei: of B. lacerta, 42 vertebra present, but their neural Siluriformes). Animal Biology 56: 39-62. prezygapophyses of the fourth vertebra is absent Esmaeili, H.R.; Teimory, A.; Owfi, F.; Abbasi, K. & Coad (Razavipour 2013). Also, the neural arch of 4th B.W. 2014a. Alien and invasive freshwater fish centrum in Weberian apparatus is bended posteriorly species in Iran: Diversity, environmental impacts in specimens of B. lacerta from the Urmia lake basin, and management. Iranian Journal of Ichthyology but in B. cyri and B. lacerta of the Tigris basin, it is 1(2): 62-72. bended dorsally. In B. lacerta, the second centrum of Esmaeili, H.R.; Coad, B.W.; Mehraban, HR., Masoudi, M., Khaefi, R., Abbasi, K., Mostavavi, H. & the caudal skeleton bears two long neural spine and Vatandoust, S. 2014b. An updated checklist of four centra form the hypural plate (Razavipour fishes of the Caspian Sea basin of Iran with a note 2013), whereas in B. cyri, the second centrum of the on their zoogeography. Iranian Journal of caudal skeleton is consumptive and the hypural plate 111 Iranian Journal of Ichthyology (June 2015), 2(2): 105-112

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