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Oreonectes Guilinensis (Teleostei, Cypriniformes, Nemacheilidae), a New Loach Species from Guangxi, China

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Oreonectes Guilinensis (Teleostei, Cypriniformes, Nemacheilidae), a New Loach Species from Guangxi, China Received: 10 August 2019 Accepted: 4 November 2019 DOI: 10.1111/jfb.14191 REGULAR PAPER FISH Oreonectes guilinensis (Teleostei, Cypriniformes, Nemacheilidae), a new loach species from Guangxi, China Jin-Qing Huang1,2 | Jian Yang3 | Zhi-Qiang Wu1 | Ya-Hui Zhao2 1College of Life Science and Technology, Guangxi University, Nanning, China Abstract 2Key Laboratory of Zoological Systematics and A new loach Oreonectes guilinensis sp. nov. is described from Guangxi Zhuang Auton- Evolution, Institute of Zoology, Chinese omous Region, China. It is the second surface-dwelling fish identified in the genus Academy of Sciences, Beijing, China 3Key Laboratory of Environment Change and Oreonectes after the type species Oreonectes platycephalus. This new species can be Resources Use in Beibu Gulf, Nanning Normal distinguished from other species within the genus by the combination of the follow- University, Nanning, China ing features: a round caudal fin, 13–14 branched rays, an incomplete lateral line with Correspondence 4–6 pores, a short pelvic fin located some distance from the anus, a scale-covered Zhi-Qiang Wu, College of Life Science and – Technology, Guangxi University, Nanning, body, a stout build (body width 14.0% 16.7% of standard length), a posterior cham- 530004,China, ber of well-developed airbladder and a yellowish-brown body with a lateral dark Email: [email protected]; Ya-Hui Zhao, Key Laboratory of Zoological Systematics and brown strip. The maximum likelihood phylogenetic analysis based on cytb gene Evolution, Institute of Zoology, Chinese recovered this new species in a well-supported clade with the type species Academy of Sciences, Beijing, 100101, China, Email: [email protected] O. platycephalus, sister to the Lefua clade. Morphological comparisons as well as our phylogenetic tree support Oreonectes shuilongensis, Oreonectes daqikongensis, Funding information This study was supported by grants Oreonectes jiarongensis as species of Troglonectes. Based on our results there are (NSFC 31972868, 31860600, and 31970382) 6 valid species in the genus Oreonectes. from the National Nature Science Foundation of China, a grant (GEFC-15-16) from National KEYWORDS Geography Society, a grant (Y229YX5105) from the Key Laboratory of the Zoological Systematics new species, Oreonectes, Pearl River, phylogeny, surface-dwelling fish and Evolution of the Chinese Academy of Sciences, a grant from the Innovation Project of Guangxi Graduate Education (T3100098001), a grant from the Education Department of Guangxi Zhuang Automous Region, China and the Guangxi Natural Science Foundation (2017GXNSFFA198010). 1 | INTRODUCTION establishing a new genus of Nemacheilidae (i.e. Oreonectes Günther 1868). Oreonectes platycephalus has several diagnostic characteristics: Nemacheilidae is the second largest family of the order Cypriniformes a flat head, a slightly compressed body and a rounded caudal fin, and is ubiquitous in diverse freshwater environments in tropical Asia, dorsal-fin origin nearer to caudal-fin base than to snout tip and poste- Europe and north-east Africa. Nemacheilids are particularly abundant in rior to pelvic-fin base, the latter of which represents an important key swift-flowing hillside streams, where they are commonly found under characteristic of the genus Oreonectes (Günther, 1868). Over the next stones (Kottelat, 2012). China, one of the countries with the richest pop- 100 years, O. platycephalus was the only species identified in this ulation of nemacheilid loaches, has been reported to harbour loaches genus but Zheng (1981) described the second species of the genus belonging to 20 genera and more than 200 species (Zhang et al., 2016). Oreonectes based on seven specimens from a cave in Guangxi, China. Based on some specimens collected in Hong Kong, Günther Another cave species Oreonectes furcocaudalis was described by Zhu (1868) described a species he named Oreonectes platycephalus, and Cao (1987), who added another diagnostic characteristic for the genus Oreonectes: a short distance between the anterior and posterior urn:lsid:zoobank.org:pub:FC23EB97-C448-4351-8DE4-F2936D36C01A. urn:lsid:zoobank.org:act:638F8391-07D5-4A09-9B1A-65CC6ACED96D. nostril, with an anterior nostril in the form of a short tube extending J Fish Biol. 2020;96:111–119. wileyonlinelibrary.com/journal/jfb © 2019 The Fisheries Society of the British Isles 111 112 FISH HUANG ET AL. into a relatively long barbel (Zhu, 1989). In the past 30 years, China. All activities followed Laboratory animal—Guideline for ethical equipped with these new diagnostic characters, dozens of cavefish review of animal welfare (GB/T 35892–2018). species were identified to belong to the genus Oreonectes. Specimens of the new species collected during field surveys in These cave-dwelling species of Oreonectes are still being discov- May 2016, in an open stream in Xingping, Guilin, Guangxi Zhuang ered and exhibit some substantial differences in terms of morphologi- Autonomous Region. Samples were collected using hand net and cal characteristics; i.e., shape of the caudal fin, position of the dorsal- mesh traps. Freshly caught fish were euthanised with eugenol. After fin origin and the caudal crests. While performing a review on the death, the pectoral fins from the right side were taken and preserved genus Oreonectes,Duet al. (2008) noticed this taxonomic problem in absolute alcohol for molecular analysis. Specimens used for mor- and tentatively divided the genus Oreonectes into two species groups; phological studies were preserved in 10% formalin, before being the platycephalus group with a rounded or truncated caudal fin, transferred to 70% ethanol for long-term storage at the collection of including Oreonectes anophthalmus Zheng 1981, O. platycephalus, the Institute of Zoology, Chinese Academy of Sciences, Beijing Oreonectes polystigmus Du, Chen & Yang 2008 and Oreonectes (ASIZB, institutional abbreviation is according to the list in Leviton retradorsalis Lan & Yang 1995 and the furcocaudalis group with a et al., 1985) and the Kunming Institute of Zoology, Chinese Academy forked caudal fin, including O. furcocaudalis and Oreonectes micro- of Sciences (KIZ). phthalmus Du, Chen & Yang 2008. Based on the original description The specimens examined (five species) for comparison belong to and drawings of Oreonectes, Kottelat (2012) suggested that some of ASIZB or KIZ. Methods used for counting and measurements were the previously identified species were not species of Oreonectes due according to Tang et al. (2012) and characteristics of cephalic lateral to their forked (v. rounded) caudal fin and their dorsal-fin originating line system were examined following guidelines from Kottelat (1990). clearly in front of the pelvic-fin origin (v. conspicuously behind the All measurements were taken point to point with dial callipers to the pelvic-fin base). These suggested species were Oreonectes elongatus nearest 0.1 mm. A few of the finer characteristics (i.e., cephalic lateral- Tang, Zhao & Zhang 2012, O. furcocaudalis, Oreonectes macrolepis Huang, line system, fins rays, nostrils distance) were examined under a micro- Du, Chen & Yang 2009, O. microphthalmus and Oreonectes translucens scope. Vertebral counts including the Weberian apparatus and the last Zhang, Zhao & Zhang 2006. Tang (2012) also came across a similar con- half-centrum were examined by micro-computed tomography (m-CT) clusion on the species within Oreonectes and suggested that a new genus images (Xradia MicroXCT-400, 30 V; Zeiss; www.zeiss.com). should be established for these species, which all possess a forked caudal A fragment of the mitochondrial gene coding for cytochrome b Š fin, a dorsal-fin origin in front of the pelvic-fin origin and well-developed (cytb) was amplified with the primers designed by lechtová et al. μ μ μ caudal crests. Subsequently, Zhang, Zhao and Tang (2016) established (2006). The 50 l PCR reactions included 5 l PCR buffer (x 10), 5 l μ μ the genus Troglonectes Zhang, Zhao & Tang 2016 with O. furcocaudalis of 2 mM deoxynucleotide triphosphate (dNTP), 2.5 lof2 M primers, 0.5 μl Taq DNA polymerase, 1.0 μl of extracted DNA and as type species, including those species with forked caudal fins and cau- 36 μlofHO. The PCR profile was 5 min at 94C, followed by dal crests, which were identified as distinct from Oreonectes (Zhang et al., 2 35 cycles of 45 s at 94C, 30 s at 54C, 1 min at 72C and ending with 2016). Meanwhile, Zhang et al. (2016) revised the genus Oreonectes as a final extension of 7 min at 72C. Amplification was visually checked having the following major diagnostic characteristics: a depressed head, on 1.0% agarose gels and PCR products were purified using a univer- nostrils separated by a short distance, anterior nostril in the form of a sal DNA clean-up kit. We obtained cytb sequences of another 23 spe- short tube that extends into a short barbel, a dorsal fin located far behind cies in the genera Oreonectes, Troglonectes, Triplophysa Rendahl 1933, the origin of the ventral and a rounded or truncated caudal fin. Currently Schistura McClelland 1838, Lefua Herzenstein 1888 and Nemacheilus there are 8 valid species in the genus Oreonectes,mostofwhichare Bleeker 1863 from GenBank (Table 1). A maximum likelihood cavefishes with narrow distributions, the only exception being the type (ML) tree building method was used to reconstruct the phylogenetic species O. platycephalus, which is the only surface-dwelling fish that is tree using MEGA 7.0, while reliability was tested using the general broadly
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