Identification of Species of Teatfish (Holothuroidea: Holothuriida) In
Total Page:16
File Type:pdf, Size:1020Kb
Plankton Benthos Res 16(3): 200–209, 2021 Plankton & Benthos Research © The Japanese Association of Benthology Identification of species of teatfish (Holothuroidea: Holothuriida) in Japan based on mitochondrial cytochrome oxidase subunit I (COI) sequences, morphology, and ossicles Iwao Tanita1,*, Shirou Nishihama2 & Takeshi Hayashibara1 1 Yaeyama Field Station, Fisheries Technology Institute, Japan Fisheries Research and Education Agency, Okinawa 907–0451, Japan 2 Fisheries Technology Institute, Japan Fisheries Research and Education Agency, Nagasaki 851–2213, Japan Received 18 November 2020; Accepted 18 January 2021 Responsible Editor: Shigeaki Kojima doi: 10.3800/pbr.16.200 Abstract: Holothurians referred to as “teatfish” contain three species, Holothuria (Microthele) nobilis, H. (M.) whitmaei and H. (M.) fuscogilva, that have recently been included in CITES Appendix II for sustainable catch and trade. Taxonomic key traits of these species are primarily genetic sequences and there are few morphological differenc- es among them, except for color patterns of the body. Although located in the northern limit of their distribution, there have been no reports identifying species of teatfish in Japan based on both morphology and genetic traits. This study investigated sequences of mitochondrial cytochrome oxidase subunit I (COI) and morphology including ossicles of black and white teatfish collected from Okinawa Island and Yaeyama Islands, Japan. The entirely black specimens and white specimens having black mottles on the dorsal side were identified as Holothuria (M.) whitmaei and H. (M.) fus- cogilva, respectively, based on the agreement of genetic and morphological traits and with published data. There is no clear difference in COI sequences between specimens from Japan and other Pacific regions for both species. The only inconsistent finding with previous reports was that one specimen of H. (M.) whitmaei lacked Cuvierian tubules, which are usually present in this species, implying plasticity of this trait. Mean diameter of the disc and height of the pillars of table ossicles were shorter and longer, respectively, in H. (M.) fuscogilva than in H. (M.) whitmaei (p<0.05, t-test). Key words: genetic, molecular, sea cucumbers, spicules include an unknown species type “pentard” (Uthicke et al. Introduction 2004, Asha 2013). These three species of teatfish in addi- Tropical holothurians are traded as luxury Chinese foods tion to elephant trunkfish H. (M.) fuscopunctata Jaeger, 1833 (“bêche-de-mer” or “trepang”) and fished all around the compose subgenus Holothuria (Microthele). Because of world, together with temperate species such as Aposticho- their high commercial value, teatfish have tended to be over- pus japonicus (Selenka, 1867). Particularly in developing exploited from an earlier period than other holothurian spe- countries of the Indo-Pacific, bêche-de-mer fisheries have cies (Eriksson & Byrne 2015), resulting in worldwide degra- been important industries for the local economy. Among dation of the resources (CITES 2019, FAO 2019). Based on the more than 58 species of edible holothurians in the world this situation, the three species H. nobilis, H. whitmaei and (Purcell et al. 2012), “teatfish” are the most commercially H. fuscogilva were included in CITES Appendix II in 2019 valuable group in tropical areas (Purcell 2014, Purcell et al. (Simone et al. 2020), which was the second listing of holo- 2018). Teatfish are composed of three species as Holothuria thurians after Isostichopus fuscus in Equador was included (Microthele) nobilis (Selenka, 1867), H. (M.) whitmaei Bell, in CITES Appendix III in 2003 (Toral-Granda 2008). Here- 1887, and H. (M.) fuscogilva Cherbonnier, 1980, and also after, species-based resource management will be required when exporting teatfish (i.e. issuing a non-detrimental find- ing: NDF). * Corresponding author: Iwao Tanita; E-mail, tanita@affrc.go.jp This CITES listing of teatfish was enabled by the recent Identification of teatfish in Japan black black and white teatfish were purchased fishermen inIslands, Yaeyama fromand two individuals of each fishermen were collectedrespectively, diving by orpurchased from on both morphological and genetic traits. vide the first description teatfish of species from Japan based three species (Uthicke This et al. 2004). study aims to pro as important evidence to (Uthicke sequences Therefore, et al. 2004). DNA were used taxonomically differentiate the and sometimes(Massin appear 1999), also in as buttons simple are occasionally lacking in to as morphological difference is that ossicles (spicules) is rudimentaryreferredAnother Uthicke et al.2004). (Massin 1999, However, fish whitmaei H. Cuvierian tubules are thick and numerous in black too variableteatfish within species to be used as taxonomic keys; still other morphological differences, which (Uthicke et al. 2004,are, Purcell 2006). There &Tekanene are however, patternsent color from the typical ones as described above cally; particularly, their young animals terns show are highly variable, both individuallyquite and ontogeni differ the their But dorsalof pat body color (Cherbonnier 1980). in most cases has irregular-shaped mottles black on the top White teatfish that is, protrusions on the lateral body (Uthicke et al. 2004). has but is also white black-colored around blotches whitmaeiH. three species was color patterns of the body; black 2004). teatfish The only clear morphological difference among the available to differentiate one from the others (Uthicke et al. toly-related each there other, are morphological few traits about the diversity this of group. in terms of local fishery management betweenand understandingthe Japanese and the known correspondspecies to, and to elucidatethe genetic relationships of teatfish, it is necessary to confirm which species Japanese teatfishabout their distribution ranges described Therefore, above. which is contraryal. 2020), to thepresent understanding Japan (Minami Minami2011, & Tamaki Yamana 2013, et the Pacific and the Indian Oceans, respectively. separated as without overlap while the distribution of the two species of black teatfish is fish ingto thelatest taxonomy(Uthicke white et al.2004), teat Accord Cherbonnier Massin 1999). 1980, (Selenka 1867, resolution (Uthicke taxonomic et al. 2004) of confusion in fuscogilvaH. Two Two and three individuals of black and white teatfish, As the three species of teatfish are phylogenetically close However, H. nobilis H. fuscogilvaH. H. fuscogilvaH. “ simplebuttons H. fuscogilvaH. and is entirely black (Bell 1887), while is entirely (Bell black 1887), and (Cherbonnier 1980), which are usually (Cherbonnier absent 1980), H. nobilis H. fuscogilvaH. Materials and Methods H.nobilis (Cherbonnier 1980, Uthicke et al. 2004). (Cherbonnier 1980, is present throughout the Indo-Pacific, H. whitmaeiH. ” are present in the ventralbody wall of sometimes has this organ, though it has been recently reported in , while often absentin whiteteat is white- orbeige-colored but H. whitmaeiH. But this. But true always is not and H. fuscogilvaH. H. nobilis H. whitmaeiH. H. nobilis “ teats in ” - - - - - - - - , Table 1. A list of collected specimens of teatfish. Sediment Specimen Date of collection Place Depth Type White teatfish (Okinawa 1) (RUMF-ZE-04001) 30th Jun 2020 Off Naha Port (Shinko Breakwater No. 1); N26°14.4′, E127°38.6′ 20–30 m Sand White teatfish (Okinawa 2) (RUMF-ZE-04002) 30th Jun 2020 Off Naha Port (Shinko Breakwater No. 1); N26°14.4′, E127°38.6′ 20–30 m Sand Black teatfish (Okinawa 3) 29th Jun 2020 Off Naha Port (Urasoe Breakwater No. 1); N26°15.6′, E127°39.3′ 15–20 m Sand-rubble, bare bedrock Black teatfish (Okinawa 4) (RUMF-ZE-04003) 4th Jul 2020 Off Naha Port (Urasoe Breakwater No. 1); N26°15.6′, E127°39.3′ 15–20 m Sand-rubble, bare bedrock White teatfish (Yaeyama 1) (RUMF-ZE-04004) 6th May 2020 Yonara Strait, Yaeyama Isls. 7–8 m Sand White teatfish (Yaeyama 2) (RUMF-ZE-04005) 14th May 2020 Yonara Strait, Yaeyama Isls. 10–20 m ̶ White teatfish (Yaeyama 3) (RUMF-ZE-04006) 20th May 2020 South of Kohama Isl., Yaeyama Isls. 12–18 m Sand-rubble Black teatfish (Yaeyama 4) (RUMF-ZE-04007) 15th May 2020 Yonara Strait, Yaeyama Isls. 5–10 m ̶ Black teatfish (Yaeyama 5) (RUMF-ZE-04008) 15th May 2020 Yonara Strait, Yaeyama Isls. 5–10 m ̶ Black teatfish (Yaeyama 6) May 2020 Yaeyama Isls. ̶ ̶ Black teatfish (Yaeyama 7) May 2020 Yaeyama Isls. ̶ ̶ Black teatfish (Yaeyama 8) ̶ Yaeyama Isls. ̶ ̶ Black teatfish (Yaeyama 9) ̶ Yaeyama Isls. ̶ ̶ 201 202 I. Tanita et al. through Urasoe-Ginowan fishery cooperative association resis in an ABI 3730xl sequencer (Applied Biosystems). (FCA) in Okinawa Island (Table 1). Black and white teat- The sequences were deposited in DNA Data Bank of Japan fish were collected from 5–20 m and 7–30 m depths, re- (DDBJ) (Table 2). spectively (Table 1). These specimens were fixed in 70% The obtained sequences were aligned using the program ethanol, after anesthetization in seawater or tapwater in ClustalW (Thompson et al. 1994) in MEGA X (Kumar et which magnesium chloride was dissolved (only for live al. 2018) together with sequences deposited in GenBank animals from Yaeyama). The fixed specimens were stored and listed in Uthicke et al. (2004) and Uthicke et al. (2010) in Ryukyu University Museum (Fujukan). (Table 2), and cut into 529 bp. Among these sequences, The number of tentacles, presence of Cuvierian tubules, AY700231.1 and AY700232.1 were eliminated from the and shapes of radial and interradial elements of calcareous analysis, because these were not located in the clade of rings were observed by dissecting the specimens. Calcare- their species in a phylogenetic tree in a pre-trial. These ous rings were observed for each three specimens of black sequences may be mixed up in the GenBank deposition and white teatfish (Okinawa 1, 2, and 4 and Yaeyama 2–5 (Dr. S. Uthicke pers. comm.). Then, a neighbor-joining tree in Table 1). (Saitou & Nei 1987) was drawn based on genetic distance To observe ossicles, small pieces of tissues were sam- calculated by Kimura two parameter (Kimura 1980) using pled from the dorsal and ventral body wall, tube feet, MEGA X.