Morphology and Phylogeny of Taeniacanthus Yamagutiishiino, 1957 (Hexanauplia: Taeniacanthidae), a Copepod Infecting the Gills Of
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J. Ocean Univ. China (Oceanic and Coastal Sea Research) https://doi.org/10.1007/s11802-020-4474-5 ISSN 1672-5182, 2020 19 (6): 1409-1420 http://www.ouc.edu.cn/xbywb/ E-mail:[email protected] Morphology and Phylogeny of Taeniacanthus yamagutii Shiino, 1957 (Hexanauplia: Taeniacanthidae), a Copepod Infecting the Gills of Rosy Goatfish Parupeneus rubescens (Mullidae) in the Arabian Gulf ABDEL-GABER Rewaida1), 2), *, AL-QURAISHY Saleh1), DKHIL Mohamed A.1), 3), ALGHAMDI Masheil1), ALGHAMDI Jawahir1), and KADRY Mohamed2) 1) Zoology Department, College of Science, King Saud University, Riyadh 11451, Saudi Arabia 2) Zoology Department, Faculty of Science, Cairo University, Cairo 12613, Egypt 3) Department of Zoology and Entomology, Faculty of Science, Helwan University, Cairo 11794, Egypt (Received February 1, 2020; revised June 18, 2020; accepted July 7, 2020) © Ocean University of China, Science Press and Springer-Verlag GmbH Germany 2020 Abstract The present study was to investigate the copepodid species infecting rosy goatfish Parupeneus rubescens, one of the most economically important fishes in the Arabian Gulf. A copepodid species identified from the examined fish specimens belongs to the Taeniacanthidae family and is labeled as Taeniacanthus yamagutii Shiino, 1957, mainly depending on its morphological, mor- phometric, and ultrastructural characteristics, in particular the presence of maxilliped claw with a conspicuous digitiform process at the base, the terminal process of the second maxilla stout, and a setiform element at the tip of each exopod spine of legs 2–4. In order to ensure the accurate identification and exact taxonomic characterization of this species, the 28S rRNA gene sequence was analyzed. The result revealed that the present copepodid species belong to the Taenicanthidae family and has a close relationship with Taeniacanthus yamagutii (gb| KR048852.1) in the same taxon. The present study demonstrated that the rosy goatfish is a host for Taeniacanthus species, which will be helpful to prevent this parasitic infection. Key words parasitic copepods; Taeniacanthidae; Taeniacanthus spp.; Arabian Gulf Sumpf, 1871; Anchistrotos Brian, 1906; Irodes Wilson, 1911; 1 Introduction Phagus Wilson, 1911; Haematophilus Wilson, 1924; Tae- niacanthodes Wilson, 1935; Parataeniacanthus Yamaguti, Taeniacanthidae is a unique family within the copepod 1939 (Accepted as Taeniacanthus Sumpf, 1871); Pseudo- order Cyclopoida, comprising members either parasitic to taeniacanthus Yamaguti and Yamasu, 1959; Echinirus Hu- marine fish or associated with sea urchins (Dojiri and Hu- mes and Cressey, 1961; Echinosocius Humes and Cressey, mes, 1982; Boxshall and Halsey, 2004). Taeniacanthids ex- 1961; Metataeniacanthus Pillai, 1963; Scolecicara Ho, 1969; hibit a high degree of host specificity at both the generic Taeniastrotos Cressey, 1969; Clavisodalis Humes, 1970; and specific levels (Boxshall and Halsey, 2004). This fa- Cirracanthus Dojiri and Cressey, 1987; Nudisodalis Do- mily, along with Bomolochidae Sumpf, 1871, Tuccidae Ver- jiri and Cressey, 1987; Biacanthus Tang and Izawa, 2005; voort, 1962, and Tegobomolochidae Avdeev, 1978, are mem- Caudacanthus Tang and Johnston, 2005; Umazuracola Ho, bers of the bomolochiform complex (Dojiri and Cressey, Ohtsuka, and Nakadachi, 2006; Saging Uyeno, Tang, and 1987; Boxshall and Halsey, 2004). They are characterized by the presence of an indistinctly four-segmented antenna; Nagasawa, 2013; Triacanthus Kim and Moon, 2013 (Ac- bearing two pectinate processes, claw-like spines, and se- cepted as Sunchenonacanthus Venmathi Maran, Moon, Ad- tae; a mandible with two subequal spinulated blades; a day, and Tang, 2016); Cepolacanthus Venmathi Maran, maxilla bearing spinulated elements; a concave ventral sur- Moon, Adday, and Tang, 2016; Suncheonacanthus Venma- face of the cephalothorax; and a lamelliform leg 1. There thi Maran, Moon, Adday, and Tang, 2016. are 25 genera with more than 91 species in Taeniacanthi- Research on ectoparasitic copepods has so far been very dae (Tang and Izawa, 2005; Tang and Johnston, 2005; Ho limited. As the number of copepodologists is relatively and Lin, 2006; Walter and Boxshall, 2019). These genera low (Ho, 2001), the morphological and anatomical fea- include Tucca Krøyer, 1837; Haemaphilus; Taeniacanthus tures are not enough to recognize and classify copepods (Hamza et al., 2007; Ramdane, 2009). Several molecular * Corresponding author. E-mail: [email protected] approaches have recently been used to check the taxo- 1410 ABDEL-GABER et al. / J. Ocean Univ. China (Oceanic and Coastal Sea Research) 2020 19: 1409-1420 nomic status of copepods independently (Fls et al., 2006; 70%, 80%, 90%, 100%) for 10 min, respectively. The sam- Ferrari and von Vaupel Klein, 2019). In copepod taxon- ples were then processed at the critical point drier ‘LEICA, omy, DNA barcoding was used to identify intra- and inter- EM CPD300’ with Freon 13, sputter-coated with gold-pal- specific morphological and molecular distinctions to help ladium in Auto fine coater (JEOL, JEC-3000FC), and fi- their phylogenetic relationship (Yazawa et al., 2008). How- nally examined and photographed under the Etec Auto- ever, very few attempts have been made to use molecular scan at 10-kV Jeol scanning electron microscope (JSM- data to restore interordinal associations within Copepoda, 6060LV) at the Molecular Biological Unit in Prince Naif as a result of the insufficient taxon sampling or sequenc- Health Research Center, King Saud University, Riyadh, ing uncertainty (Kim and Kim, 2000; Ferrari et al., 2010). Saudi Arabia. Huys et al. (2007) found that nuclear ribosomal genes The different body parts of the recovered copepods were 18S and 28S rRNA include semi-conserved domains that measured using the Olympus ocular micrometer and the intersperse with divergent regions, allowing for a wide average values with the range given in parentheses were range of taxonomic levels of phylogenetic reconstruction. employed for analysis. Sewell’s style (1949) was adopted There is a need for more extensive work to gain a better for the armature formula of the swimming legs, in which understanding of the parasitic copepods infecting the Ara- the spines and setae are denoted by Roman and Arabic bian Gulf fish in general and those off Saudi Arabia in numerals, respectively. particular. The purpose of this research is to provide com- plete data on parasitic copepods and their indices in the 2.3 Molecular Analysis rosy goatfish Parupeneus rubescens from the Arabian Gulf 2.3.1 DNA extraction and polymerase chain in Saudi Arabia. In addition, partial sequences of 28S rRNA reaction (PCR) gene have been developed and used to evaluate the phy- Genomic DNA was extracted from ethanol-contained logenetic status of this species within Taeniacanthidae. samples using the DNeasy tissue kit© (Qiagen, Hilden, Germany) following the manufacturer’s instructions. DNA 2 Materials and Methods quality and purity were quantified with the NanoDrop ND-1000 spectophotometer (Thermo Fischer Scientific, 2.1 Fish Collection Inc., Wilmington, DE, USA) and 20 ng of genomic DNA A total of 80 specimens of the rosy goatfish, Paru- was used for PCR amplification. The 28S rRNA gene re- peneus rubescens (F: Mullidae), were collected from land- gion was amplified by PCR and subsequently sequenced. ing sites on the coasts of the Arabian Gulf off Dammam The PCR for this region was produced in a total of 20 µL City in Saudi Arabia. They were immediately transported of reaction volume containing 4 μL 5× FIREPol® Master to the Laboratory of Parasitology Research, Zoology De- Mix (Solis BioDyne), 2 μL of genomic DNA, 0.6 μL of partment, College of Science, King Saud University, Ri- each primer, and completed to the required volume of 20 yadh, Saudi Arabia. All procedures comply with the ethi- µL by nuclease-free water. PCR amplification was per- cal standards approved by the Institutional Review Board formed using the following primers: 28SF, 5’-ACA ACT (IRB) of King Saud University, Riyadh, Saudi Arabia. Sam- GTG ATG CCC TTA G-3’ and 28SR, 5’-TGG TCC GTG ples of fish were classified according to the standards of TTT CAA GAC G-3’ previously designed by Song et al. the database of fishbase.org. They were thoroughly check- (2008). The amplification technique of the PCR thermo- ed for the presence of parasitic crustacean infections on cycler profile and the primer combination used to amplify their body surface, fins, head, gill filaments, oral cavities this genetic marker was done according to Song et al. and other tissues, based on the method described previ- (2008). All PCR products were verified on 1% agarose gel ously by Ravichandran et al. (2007). in 1× Tris-acetate-EDTA (TAE) fixed with 1% ethidium bromide and then visualized with a UV trans-illuminator. The PCR products of the expected size were gel-excised, 2.2 Parasitological Studies purified and cloned using the manufacturer’s instructions 2.2.1 Light microscopic examination for the PureLinkTM Quick Gel Extraction Kit (Qiagen, Parasitic copepods were removed using a special nee- Hilden, Germany). dle. They were washed several times in a saline solution, preserved in 70% ethyl alcohol, dehydrated in a 2-h glyc- 2.3.2 Sequence alignment and phylogenetic analysis erin ethanol series, and mounted as temporary preparations Sequencing was performed on 3130×l Genetic Analy- in lactophenol as described by Pritchard and Kruse (1982). zer (Biosystems® 3130, Thermo Fisher Scientific, USA) Prepared samples were analyzed and photographed