Checklist and DNA Barcoding of the Scorpaenidae (Teleostei: Scorpaeniformes) in Taiwan

© 2024 Academia Sinica, Taiwan Open Access Checklist and DNA Barcoding of the Scorpaenidae (Teleostei: Scorpaeniformes) in Taiwan Tak-Kei Chou1, Wen-Chien Huang1, Wei-Cheng Jhuang2,3 , and Te-Yu Liao1,* 1Department of Oceanography, National Sun Yat-sen University, Kaohsiung, Taiwan. *Correspondence: swp01[email protected] (Liao) E-mail: [email protected] (Chou); [email protected] (Huang) 2Doctoral Degree Program in Marine Biotechnology, National Sun Yat-sen University, Kaohsiung, Taiwan. E-mail: fish101[email protected] (Jhuang) 3Doctoral Degree Program in Marine Biotechnology, Academia Sinica, Taipei, Taiwan Received 19 January 2024 / Accepted 26 June 2024 / Published 25 December 2024 Communicated by Chien-Hsiang Lin Species of the family Scorpaenidae are easily misidentified due to their similar appearances, a result of camouflaging to their surroundings. In recent years, many species from this family have been described, and generic placements of some species have been revised. Previously, there were 80 species belonging to 29 genera of the Scorpaenidae recorded in Taiwanese waters. However, their taxonomy has not been revised for decades. It is necessary to update the checklist of the Scorpaenidae occurring in Taiwanese waters based on updated morphological and molecular data. In the present study, we revised the Taiwanese scorpaenids based on 296 specimens and updated the checklist, amounting to a total of 85 species of 29 genera, of which Sebastapistes mauritiana (Cuvier) is a new record, and three species from the genera Phenacoscorpius, Scorpaenopsis, and Sebastapistes are unable to be identified to any species. Using molecular analysis, we conducted the first comprehensive DNA barcoding study of the Scorpaenidae from Taiwanese waters based on a partial cytochrome c oxidase I (COI) gene of 655 bps. A total of 118 COI sequences were generated from voucher specimens of 66 species (28 genera) identified based on morphological characters. The COI sequences of Parascorpaena maculipinnis, Scorpaena pepo, and Scorpaenopsis orientalis are new to online databases. According to the Kimura-2 Parameter (K2P) genetic distance, the mean interspecific variation (15.61%) was distinctly greater than the mean intraspecific variation (0.22%), suggesting a barcoding gap. The maximum likelihood tree showed that all lineages were supported by high bootstrap values. Key words: Diversity, Ichthyofauna, Mitochondrion, Stonefish, Taxonomy BACKGROUND Members of the family Scorpaenidae are known for the venomous spines on their dorsal, pelvic, and anal fins (Allen and Eschmeyer 1973; Nelson 2006), and their ability to camouflage with their surroundings using variable color patterns, tentacles, flaps, and barbels (Poss 1999; Randall and Eschmeyer 2002; Krzyżak and Korzeniewski 2021). This family is distributed in a variety of habitats, such as intertidal and sublittoral zones, sandy substrates, coral reefs, rocky reefs, and continental shelves with a depth range from 0 to 1,600 m (Masuda et al. 1984; Fedorov et al. 2003; Poss and Eschmeyer 2003; Randall 2005a). The Scorpaenidae are a species-rich family, composed of 36 genera and more than 350 valid species (Nelson et al. 2016; Fricke et al. 2024), but their definition has been contentious (Matsubara 1943; Washington et al. 1984; Ishida 1994; Poss 1999; Imamura 2004; Shinohara and Imamura 2005; Smith Citation: Chou TK, Huang WC, Jhuang WC, Liao TY. 2024. Checklist and DNA barcoding of the Scorpaenidae (Teleostei: Scorpaeniformes) in Taiwan. Zool Stud 63:37. doi:10.6620/ZS.2024.63-37. doi:10.6620/ZS.2024.63-37. Zoological Studies 63:37 (2024) doi:10.6620/ZS.2024.63-37 1 © 2024 Academia Sinica, Taiwan et al. 2018). In the present study, the definition of the Scorpaenidae follows the classification of Smith et al. (2018) which was based on morphological and molecular characteristics of a large number of Scorpaeniformes specimens. Poss (1999) and Smith et al. (2018) proposed that this family is characterized by the presence of a suborbital stay firmly connected to the preopercle, spines on head, gill membranes free from isthmus, compressed body, presence of scales, 24 to 30 vertebrae, venomous glands of the dorsal, anal, and pelvic fin spines, dorsal fin with XII to XIII spines (rarely VIII), anal fin with III spines (rarely II), pelvic fins with one spine and five rays, and a well-developed pectoral fin with lower rays unbranched. Morphological identification of scorpaenids has been considered difficult because of limited diagnostic characters and variable color patterns based on surroundings for most species (Poss 1999; Randall and Eschmeyer 2002; Randall 2005a b; Krzyżak and Korzeniewski 2021). In the last two decades, most taxonomic work on Scorpaenidae was conducted based on specimens from the western Pacific, including descriptions of new species and/or synonymizations of nominal species in the genera Brachypterois, Dendrochirus, Pterois, Scorpaena, Scorpaenopsis, Sebastapistes, Sebastiscus, Parascorpaena, Lythrichthys, Neomerinthe, Phenacoscorpius, Pteroidichthys, Scorpaena, and Scorpaenodes (Randall and Eschmeyer 2002; Allen and Erdmann 2008; Motomura 2008 2009; Motomura and Senou 2008; Motomura et al. 2010a b 2014 2015 2016; Matsunuma et al. 2013 2017; Motomura and Kanade 2015; Matsunuma and Motomura 2015 2019; Morishita et al. 2018; Wibowo and Motomura 2019a b; Hoshino and Motomura 2021; Wada et al. 2021; Chou and Liao 2022; Chou et al. 2023; Matsumoto et al. 2023; Matsumoto and Motomura 2024). Among these studies, only Matsunuma et al. (2017), Wada et al. (2021), Chou and Liao (2022), and Chou et al. (2023) provided molecular characters of scorpaenids. In Taiwan, the first study on the diversity of the Scorpaenidae was Chen (1969)’s synopsis of the vertebrates of Taiwan, in which 13 species from nine genera were recorded. Later, Chen (1981) revised the taxonomy of the Scorpaenidae from Taiwan and recorded 42 species of 25 genera, and Shen et al. (1993) updated the list to 48 species of 26 genera. Recent taxonomic studies of the Scorpaenidae have greatly expanded the biodiversity of this fish group in Taiwan (Randall and Eschmeyer 2002; Chen 2003; Motomura et al. 2007 2009a b 2010a 2011; Shao et al. 2008; Motomura and Senou 2009; Chen et al. 2010; Shen and Wu 2011; Morishita et al. 2018; Koeda and Ho 2019; Koeda et al. 2019; Chou 2021; Chou and Tang 2021; Wada et al. 2021; Chou and Liao 2022), amounting to a total of 80 species placed in 29 genera. Despite the increased number of recorded species, there was no taxonomic revision of the Scorpaenidae made from Taiwanese waters after Chen (1981). Furthermore, generic placements of some species have changed (Randall and Poss 2002; Motomura et al. 2010b; Poss et al. 2010; Wada et al. 2021), and some species in the western Pacific were considered junior synonyms or misidentifications (Nakabo 2002; Motomura et al. 2009a b 2010b 2015; Wibowo and Motomura 2019a b; Hoshino and Motomura 2021; Wada et al. 2021). These recent taxonomic works imply that the checklist of scorpaenids in Taiwan needs revision based on museum collections and recent taxonomic literature. In addition, DNA barcoding is needed to provide molecular references for the scorpaenids of Taiwan. In the present study, we aimed to: (a) provide a reliable DNA barcoding reference of the Scorpaenidae from Taiwanese waters based on morphological examination; and (b) update the checklist of the Scorpaenidae present in Taiwan. MATERIALS AND METHODS Sampling and morphological analyses Fresh specimens were collected in Taiwanese waters from 2017 to 2022 using hand nets and angling, as well as purchasing from local fish markets. Several islands around Taiwan were also surveyed, including Penghu, Liuqiu, Lanyu (Orchid Island), and Green Islands (Fig. 1). Fresh specimens were fixed in 10% neutral buffered formalin and preserved in 70% ethanol thereafter. Some examined materials were loaned from museum collections of the Academia Sinica Institute of Zoology, Taipei (ASIZP), and the National Museum of Marine Biology and Aquarium (NMMB-P), Pingtung. At least five specimens per species were examined whenever possible. The definition of habitat types in Taiwan followed Shao et al. (2008). Terminology and definitions of morphometrics and meristics generally followed Motomura (2004a b), Motomura et al. (2005a– c), and Motomura and Johnson (2006). The terminology of head spines followed Randall and Eschmeyer (2002). The meristics were examined on the left side of fish. The last two rays of the dorsal and anal fins were counted as one. The vertebra count was determined by X-radiographs. Measurements were carried out using a digital caliper with 1 mm precision. At least two tissue samples per species (when possible) were taken from the fin clips or dorsal muscle and preserved in 95% ethanol. Some tissue samples were loaned from the cryobank of the Research Center for Biodiversity page 2 of 18Zoological Studies 63:37 (2024) © 2024 Academia Sinica, Taiwan of Academia Sinica. Voucher specimens collected in this study were deposited in ASIZP, Department of Oceanography, National Sun Yat-sen University, Kaohsiung (DOS), and NMMB-P. DNA extraction, amplification, and sequencing DNA was extracted from tissue samples using GeneMark Easy Tissue & Cell Genomic DNA Purification Kit following the manufacturer’s protocol. The polymerase chain reaction (PCR) was used to amplify the cytochrome c oxidase subunit I (COI) gene. PCR products were amplified in a 25 μL volume containing 3 μL of 10X Taq Buffer, 2 μL of dNTP mixture at 10 mM, 1 μL of forward and reverse primers at 5 μM, 0.13 μL of Pro Taq Plus DNA polymerase (Protech Technology Enterprise, Taiwan), 1 μL of template DNA, and 16.87 μL of ultrapure water. The COI fragment was amplified using Ward et al.’s (2005) universal COI primers, and ScorF (5'- CTCAGCCATCCTACCTGTGG-3') and ScorR (5'- ACTTCTGGGTGRCCGAAGAA-3') designated by the present study. The thermal PCR condition of COI was composed of an initial denaturation step at 95°C for 4 min, then 35 cycles of 94°C for 30 s, 48°C for 30 s and 72°C for 1 min, and a final extension at 72°C for 10 min. PCR products were visualized on 2% agarose gels and subsequently purified using SAP-Exo Kit (Jena Bioscience). PCR products were sequenced in both forward and reverse directions by a biotechnology company (Genomics, Taiwan). The forward and reverse sequences were edited and assembled using BioEdit ver. 7.2.5 (Hall 1999). All sequences were deposited in Genbank (Table S1). Sequence analysis DNA sequences were aligned using ClustalW (Thompson et al. 1994) in BioEdit ver. 7.2.5 (Hall 1999). The substitution saturation of COI mutation was tested using DAMBE ver. 7.0.10 (Xia 2018). The Kimura-two parameter (K2P) model was implemented for COI gene in phylogenetic reconstruction and distance metrics among species. Phylogenetic analysis of COI sequences was conducted with the Maximum likelihood (ML) method using MEGA ver. 10.1.1 (Kumar et al. 2018). Branch support value was assessed using the bootstrapping criterion with 1,000 replicates. Synanceia verrucosa (accession number: JQ432179) of the Synanceiidae was chosen as outgroup for phylogenetic analysis. Genetic divergences at different taxonomic levels (inter-generic, inter-specific, intraspecific) were calculated using MEGA ver. 10.1.1 (Kumar et al. 2018). All COI sequences were compared with those from public databases using Basic Local Alignment Search Tool (BLAST) on GenBank and the BOLD Identification System (IDS) (Ratnasingham and Hebert 2007; Johnson et al. 2008). Sequences with similarity values greater than 98% were considered to be conspecific (Huang et al. 2023). Ethics statement The study was conducted in strict accordance with the Wildlife Conservation Act in Taiwan. No ethical approval was required for this study since all species in this study were not protected species and not listed in CITES. Some specimens were collected from Liuqiu and Kenting with the approvals of the Dapeng Bay National Scenic Area Administration, Tourism Bureau (Project #NAMR110029; Collection Permit #11005508700) and the Kenting National Park Headquarters (Project #NAMR110029; Collection Permit #1091002868), respectively. All individuals were not involved in animal experiments. RESULTS A total of 296 scorpaenid specimens belonging to 85 species from 29 genera (Table 1) were examined and literature reviewed in the present study (Table S1, Fig. S1), of which one species, Sebastapistes mauritiana Fig. 1. Map of sampling sites in this study. page 3 of 18Zoological Studies 63:37 (2024) © 2024 Academia Sinica, Taiwan Table 1. Checklist of the Scorpaenidae from Taiwanese waters Brachypterois Brachypterois serrulata (Richardson, 1846)/ CS; m1: Brachypterois serrulatus Brachypterois serrulifer Fowler, 1938/ CS Caracanthus Caracanthus maculatus (Gray, 1831)/ SR Caracanthus unipinna (Gray, 1831)/ SR Dendrochirus Dendrochirus zebra (Cuvier, 1829)/ SR Ebosia Ebosia bleekeri (Döderlein, 1884)/ CS Ectreposebastes Ectreposebastes imus Garman, 1899/ DCS Helicolenus Helicolenus hilgendorfii (Döderlein, 1884)/ DCS; m1: Helicolenus hilgendorfi Hoplosebastes Hoplosebastes armatus Schmidt, 1929/ DCS Iracundus Iracundus signifer Jordan & Evermann, 1903/ SR Lythrichthys Lythrichthys cypho (Fowler, 1938)/ DCS Lythrichthys eulabes Jordan & Starks, 1904/ DCS Lythrichthys longimanus (Alcock, 1894)/ DCS; c: Setarches longimanus Nemapterois Nemapterois biocellatus Fowler, 1938/ SR; c: Dendrochirus biocellatus Neochirus Neochirus bella (Jordan & Hubbs, 1925)/ CS; c: Brachirus bellus, Dendrochirus bellus Neochirus brachyptera (Cuvier, 1829)/ SR; c: Dendrochirus brachypterus Neomerinthe Neomerinthe erostris (Alcock, 1896)/ DCS; s: Neomerinthe rotunda Neomerinthe ignea Matsumoto, Muto & Motomura, 2023/ DCS Neomerinthe kaufmani (Herre, 1952)/ DCS Neomerinthe megalepis (Fowler, 1938)/ DCS Neomerinthe procurva Chen, 1981/ DCS Parapterois Parapterois heterurus (Bleeker, 1856)/ SR; m1: Parapterois heterura Parascorpaena Parascorpaena aurita (Rüppell, 1838)/ SR; m2: Parascorpaena picta, s: Scorpaena bynoensis Parascorpaena maculipinnis Smith, 1957/ SR Parascorpaena mcadamsi (Fowler, 1938)/ SR Parascorpaena mossambica (Peters, 1855)/ SR Parascorpaena poseidon Chou & Liao, 2022/ SR Phenacoscorpius Phenacoscorpius megalops Fowler, 1938/ DCS Phenacoscorpius sp./ DCS Pontinus Pontinus macrocephalus (Sauvage, 1882)/ DCS Pontinus tentacularis (Fowler, 1938)/ DCS Pteroidichthys Pteroidichthys acutus Motomura & Kanade, 2015/ CS Pteroidichthys amboinensis Bleeker, 1856/ CS Pteroidichthys noronhai (Fowler, 1938)/ CS; c: Pteropelor noronhai Pterois Pterois lunulata Temminck & Schlegel, 1843/ SR Pterois russelii Bennett, 1831/ SR; m1: Pterois russelli, Pterois russellii Pterois volitans (Linnaeus, 1758)/ SR Pteropterus Pteropterus antennatus (Bloch, 1787)/ SR; c: Pterois antennata Pteropterus paucispinula (Matsunuma & Motomura, 2014)/ SR; c: Pterois paucispinula Pteropterus radiatus (Cuvier, 1829)/ SR; c: Pterois radiata page 4 of 18Zoological Studies 63:37 (2024) © 2024 Academia Sinica, Taiwan Rhinopias Rhinopias eschmeyeri Condé, 1977/ SR Rhinopias frondosa (Günther, 1892)/ SR; m2: Rhinopias aphanes Scorpaena Scorpaena miostoma Günther, 1877/ CS; m2: Parascorpaena picta Scorpaena neglecta Temminck & Schlegel, 1843/ CS; s: Scorpaena izensis Scorpaena onaria Jordan & Snyder, 1900/ CS Scorpaena pepo Motomura, Poss & Shao, 2007/ CS; m2: Parascorpaena picta Scorpaenodes Scorpaenodes albaiensis (Evermann & Seale, 1907)/ SR Scorpaenodes evides (Jordan & Thompson, 1914)/ SR; s: Scorpaenodes littoralis Scorpaenodes guamensis (Quoy & Gaimard, 1824)/ SR Scorpaenodes hirsutus (Smith, 1957)/ SR Scorpaenodes kelloggi (Jenkins, 1903)/ SR Scorpaenodes minor (Smith, 1958)/ SR Scorpaenodes parvipinnis (Garrett, 1864)/ SR Scorpaenodes quadrispinosus Greenfield & Matsuura, 2002/ SR Scorpaenodes scaber (Ramsay & Ogilby, 1886)/ SR Scorpaenodes varipinnis Smith, 1957/ SR Scorpaenopsis Scorpaenopsis cirrosa (Thunberg, 1793)/ SR; m1: Scorpaenopsis cirrhosa Scorpaenopsis cotticeps Fowler, 1938/ SR Scorpaenopsis diabolus (Cuvier, 1829)/ SR; m1: Scorpaenopsis diabolis Scorpaenopsis macrochir Ogilby, 1910/ SR Scorpaenopsis neglecta Heckel, 1837/ SR; m2: Scorpaenopsis gibbosa Scorpaenopsis obtusa Randall & Eschmeyer, 2002/ SR Scorpaenopsis orientalis Randall & Eschmeyer, 2002/ SR Scorpaenopsis oxycephala (Bleeker, 1849)/ SR Scorpaenopsis papuensis (Cuvier, 1829)/ SR Scorpaenopsis possi Randall & Eschmeyer, 2002/ SR Scorpaenopsis ramaraoi Randall & Eschmeyer, 2002/ SR Scorpaenopsis sp./ SR Scorpaenopsis venosa (Cuvier, 1829)/ SR Scorpaenopsis vittapinna Randall & Eschmeyer, 2002/ SR Sebastapistes Sebastapistes cyanostigma (Bleeker, 1856)/ SR; s: Scorpaena albobrunnea Sebastapistes fowleri (Pietschmann, 1934)/ SR Sebastapistes mauritiana (Cuvier, 1829)*/ SR Sebastapistes sp./ SR Sebastapistes strongia (Cuvier, 1829)/ SR; s: Sebastapistes kowiensis Sebastapistes tinkhami (Fowler, 1946)/ SR Sebastes Sebastes joyneri Günther, 1878/ CS Sebastes thompsoni Jordan & Hubbs, 1925/ CS Sebastiscus Sebastiscus albofasciatus (Lacepède, 1802)/ DCS; c: Sebastes albofasciatus Sebastiscus marmoratus (Cuvier, 1829)/ DCS; c: Sebastes marmoratus Sebastiscus tertius (Barsukov & Chen, 1978)/ DCS; c: Sebastes tertius Sebastiscus vibrantus Morishita, Kawai & Motomura, 2018/ DCS Setarches Setarches guentheri Johnson, 1862/ DCS Taenianotus Taenianotus triacanthus Lacepède, 1802/ SR Thysanichthys Thysanichthys crossotus Jordan & Starks, 1904/ DCS Asterisks (*) represent new records. Abbreviations: Habitat type— CS (coastal shore); SR (shallow reef, less than 60 m); DCS (deep continental shelf, deeper than 100 m depth). s - synonym; c - formerly used combination; m1 - misspelling; m2 - misidentification. Table 1. (Continued) page 5 of 18Zoological Studies 63:37 (2024) © 2024 Academia Sinica, Taiwan (Cuvier, 1829), was newly recorded, and three species of Phenacoscorpius, Scorpaenopsis, and Sebastapistes were not able to be identified to any known species. In total, 118 COI sequences belonging to 66 species of 28 genera were generated, in which sequences of Parascorpaena maculipinnis, Scorpaena pepo, and Scorpaenopsis orientalis were new to online databases (GenBank and BOLD systems). Eight species in four genera were only examined morphologically without molecular data (Table S1). DNA barcoding All data are available in GenBank with accession numbers and catalog numbers of voucher specimens listed in table S1. After alignment, the consensus length of all COI fragments was 655 bps. The saturation was tested for the entire fragment and each codon position of the COI sequences using DAMBE v. 7.0.10 (Xia 2018), and no signs of saturation were detected. No insertion, deletion, and stop codon were found. The ML tree is shown in figure 2. All morphologybased species were monophyletic groups supported by high bootstrap values. Three genera of the Scorpaenidae were non-monophyletic, including Pteroidichthys, Scorpaenodes, and Sebastapistes. The pairwise genetic distances at different taxonomic levels are summarized in table 2. The intraspecific K2P distance was between 0 and 1.37%, with a mean of 0.22%. The maximum value was found in Helicolenus hilgendorfii (1.37%). The interspecific K2P distance was from 0.46 to 32.32% with a mean of 15.61%. Several species pairs had interspecific distances lower than 2%, including Neochirus bella vs. Neochirus brachyptera (0.69%), Pterois lunulata vs. Pterois russelii (0.62%), Pterois lunulata vs. Pterois volitans (0.85%), Pterois russelii vs. Pterois volitans (0.85%), and Sebastiscus tertius vs. Sebastiscus vibrantus (0.46%). Overall, the mean interspecific distances were over 70-fold higher than the mean intraspecific distances. Aside from the abovementioned five species pairs, the distribution of genetic distances (Fig. 3) also showed a barcoding gap between intraspecific and interspecific divergences. After blasting in GenBank and BOLD databases, sequences of 22 species of nine genera were found to have more than one species with similarities ≥ 98% (Table S2). Three specimens of three genera, Phenacoscorpius, Scorpaenopsis and Sebastapistes, were unsuccessfully identified to any species and their sequences do not match any species in the online database. The Scorpaenidae fauna in Taiwanese waters The updated checklist of the Scorpaenidae from Taiwanese waters is shown in table 1. The counts of dorsal, pectoral, and anal fins of all examined specimens are shown in table 3, while the standard length of all examined specimens is shown in table S1. In the present study, 11 species listed in the checklist have a lack of examined specimens and sequences, as their records are only based upon references. Remarks on the new records, species without examined specimens, taxonomically uncertain species, and species pairs with low genetic distance were provided as follows. Iracundus signifer Jordan & Evermann, 1903 Remarks: No specimen was examined in this study. The record was based on a specimen (BPBMI23411) collected in Nanwan, Pingtung by J.E. Randall in 1978 (Chen 1981), in which some specimens were collected from Taiwanese waters. Lythrichthys cypho (Fowler, 1938) Remarks: No specimen was examined in this study. The record was based on Wada et al. (2021). Lythrichthys longimanus (Alcock, 1894) Remarks: No specimen was examined in this Table 2. Summary of K2P genetic distances at different taxonomic levels Taxonomic level Distance (%) Minimum Maximum Mean ± SE Intra-specific (52 species) 0.00 1.40 0.22 ± 0.04 Inter-specific (41 species) 0.46 32.32 15.61 ± 0.58 Inter-generic (24 genera) 7.19 27.77 21.19 ± 0.19 SE, standard error. Species with only one sequence were excluded from intra-specific distance calculations; genera with only one species were excluded from inter-specific distance calculations within the same genus; families with only one genus were excluded from inter-genus distance calculations within the same family. page 6 of 18Zoological Studies 63:37 (2024) © 2024 Academia Sinica, Taiwan Fig. 2. The maximum likelihood tree based on 118 COI sequences of 66 species of the Scorpaenidae collected from Taiwanese waters. Numerals on nodes represent bootstrap values. Bootstrap values below 70 are not shown. Right hand side labels mark non-monophyletic genera. Color blocks on lineages denote the taxonomically uncertain taxa, including Phenacoscorpius sp. (yellow), Scorpaenopsis sp. (blue), and Sebastapistes sp. (red). page 7 of 18Zoological Studies 63:37 (2024) © 2024 Academia Sinica, Taiwan study. The record was based on Wada et al. (2021), in which some specimens were collected from Taiwanese waters. Neochirus bella (Jordan & Hubbs, 1925) Remarks: This species was proposed as a member of the Dendrochirus (= Neochirus) brachypterus complex due to the similarity in overall body appearance (Matsunuma et al. 2017). Neochirus bella could be distinguished from N. brachyptera by the lower count of longitudinal scale series (ca. 34 in N. bella vs. 45–54 in N. brachyptera). Based on our examined materials, the lower count of longitudinal scale series (29–33) matched the description of N. bella. Neochirus brachyptera (Cuvier, 1829) Remarks: The comparison between Neochirus bella and N. brachyptera was shown in the remark of Neochirus bella. Based on our examined materials, the higher count of longitudinal scale series (41–44) matched the description of N. brachyptera. Neomerinthe ignea Matsumoto, Muto & Motomura, 2023 Remarks: No specimen was examined in this study. The record was based on Matsumoto et al. (2023), in which some specimens were collected from Taiwanese waters. Neomerinthe megalepis (Fowler, 1938) Remarks: No specimen was examined in this study. The first record was reported by Chen (1981) based on six specimens (CAS27744, SDSC73-37, SIO80-206, SIO80-207, SIO80-208, SIO80-221) from Tungkang, Pingtung. But these specimens showed more scales in longitudinal series (36–41), a discrepancy from the 25–30 scales in the original description (Fowler 1938; Herre 1952). These specimens are actually N. kaufmani based on the number of longitudinal series. We also examined a specimen (ASIZP0064297) originally identified as N. megalepis and found that it is a misidentification of N. kaufmani. However, some specimens of N. megalepis were collected from Taiwanese waters by Matsumoto and Motomura (2024). Fig. 3. Distribution frequency of K2P genetic distances (%) for COI of the Scorpaenidae at different taxonomic levels. Table 3. Frequency distribution of spine and ray counts on dorsal, pectoral, and anal fins in Taiwanese species of the Scorpaenidae Dorsal-fin spines Dorsal-fin rays Pectoral-fin rays Anal-fin spines Anal-fin rays 7 8 / 11 12 13 7 8 9 10 11 12 13 14 12 13 14 15 16 17 18 19 20 21 22 2 3 4 5 6 7 / 11 12 13 14 15 Brachypterois serrulatus 5 4 1 1 4 5 5 Brachypterois serrulifer 3 2 1 3 2 3 Caracanthus maculatus 23 311 113 5 211 Caracanthus unipinna 1 1 1 1 1 Dendrochirus biocellatus 5 5 2 3 5 5 Dendrochirus zebra 1 4 5 4 1 5 5 Ebosia bleekeri 2 3 1 3 1 4 5 1 3 1 Ectreposebastes imus 5 1 1 3 1 2 1 1 5 1 4 Helicolenus hilgendorfii 5 2 3 4 1 5 5 Hoplosebastes armatus 5 3 2 1 2 2 5 5 Lythrichthys eulabes 5 1 4 5 5 5 page 8 of 18Zoological Studies 63:37 (2024) © 2024 Academia Sinica, Taiwan Dorsal-fin spines Dorsal-fin rays Pectoral-fin rays Anal-fin spines Anal-fin rays 7 8 / 11 12 13 7 8 9 10 11 12 13 14 12 13 14 15 16 17 18 19 20 21 22 2 3 4 5 6 7 / 11 12 13 14 15 Neochirus bella 5 2 3 2 3 5 5 Neochirus brachyptera 2 2 2 2 2 Neomerinthe erostris 1 4 4 1 5 5 3 2 Neomerinthe kaufmani 5 5 1 3 1 5 5 Neomerinthe procurva 4 4 4 4 2 2 Parapterois heterura 2 3 2 3 1 3 1 5 1 4 Parascorpaena aurita 5 1 4 5 5 5 Parascorpaena maculipinnis 5 5 1 4 5 5 Parascorpaena mcadamsi 5 5 1 4 5 5 Parascorpaena mossambica 5 5 1 4 5 5 Parascorpaena poseidon 5 5 4 1 5 5 Phenacoscorpius sp. 1 1 1 1 1 Pontinus tentacularis 5 5 5 1 4 5 Pteroidichthys acutus 1 1 1 1 1 Pteroidichthys amboinensis 3 3 1 1 1 3 3 Pteroidichthys noronhai 1 1 1 1 1 Pterois lunulata 1 1 1 1 1 Pterois paucispinula 5 5 4 1 5 5 Pterois russelii 5 1 4 1 4 5 1 4 Pterois volitans 5 1 4 5 5 1 4 Pteropterus antennatus 3 2 1 4 3 1 1 5 5 Pteropterus radiatus 5 2 3 3 2 5 5 Rhinopias eschmeyeri 3 3 1 2 3 2 1 Rhinopias frondosa 3 3 1 2 3 3 Scorpaena miostoma 5 5 5 5 5 Scorpaena neglecta 5 5 4 1 5 5 Scorpaena onaria 2 2 2 2 2 Scorpaena pepo 5 5 5 5 5 Scorpaenodes albaiensis 5 1 3 1 1 4 5 1 4 Scorpaenodes evides 5 5 4 1 5 5 Scorpaenodes guamensis 5 5 1 4 5 5 Scorpaenodes kelloggi 5 2 3 2 3 5 5 Scorpaenodes minor 2 2 2 2 2 Scorpaenodes parvipinnis 5 5 4 1 5 5 Scorpaenodes scaber 1 1 1 1 1 Scorpaenodes varipinnis 5 3 2 4 1 5 5 Scorpaenopsis cirrosa 5 1 4 2 3 5 5 Scorpaenopsis cotticeps 2 2 2 2 2 Scorpaenopsis diabolus 5 5 5 5 5 Scorpaenopsis macrochir 3 1 2 3 3 3 Scorpaenopsis neglecta 5 5 3 2 5 5 Scorpaenopsis orientalis 5 5 5 5 5 Scorpaenopsis oxycephala 5 5 1 4 5 5 Scorpaenopsis papuensis 5 5 1 4 5 5 Scorpaenopsis possi 5 5 5 5 5 Scorpaenopsis ramaraoi 5 5 1 4 5 5 Scorpaenopsis sp. 1 1 1 1 1 Scorpaenopsis venosa 5 4 1 5 5 5 Scorpaenopsis vittapinna 2 2 2 2 2 Sebastapistes cyanostigma 5 5 1 4 5 5 Sebastapistes fowleri 5 1 4 1 4 5 5 Sebastapistes mauritiana 1 1 1 1 1 Sebastapistes sp. 1 1 1 1 1 Sebastapistes strongia 5 5 1 4 5 5 Sebastapistes tinkhami 3 3 3 3 3 Sebastes thompsoni 1 1 1 1 1 Sebastiscus albofasciatus 5 5 4 1 5 5 Sebastiscus marmoratus 5 5 1 4 5 5 Sebastiscus tertius 5 1 4 2 3 5 4 1 Sebastiscus vibrantus 2 1 1 2 2 2 Setarches guentheri 2 1 1 1 1 2 2 Taenianotus triacanthus 41 122 41 5 131 Thysanichthys crossotus 5 1 4 1 4 5 5 Table 3. 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Following the scientific name are the catalog number, standard length (SL), and GenBank accession number of the specimen. NA for the accession number indicates that the specimen was used solely for morphological examination and was not sequenced. (download) Table S1. List of species, body size, catalog number of specimens, and their accession numbers. Species without examined specimen are denoted by a hyphen (-) representing data as “not available” . (download) Table S2. List of morphological and molecular identifications. The listed sequences included more than one species after BLAST. (download) page 18 of 18Zoological Studies 63:37 (2024)