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(Cyprinidae: Labeoninae ) Originating from Bangka and Belitung Islands, Indonesia

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(Cyprinidae: Labeoninae ) Originating from Bangka and Belitung Islands, Indonesia BIODIVERSITAS ISSN: 1412-033X Volume 22, Number 2, February 2021 E-ISSN: 2085-4722 Pages: 794-802 DOI: 10.13057/biodiv/d220233 First genetic record and the phylogenetic relationship of Osteochilus spilurus (Cyprinidae: Labeoninae ) originating from Bangka and Belitung Islands, Indonesia ARDIANSYAH KURNIAWAN1,2,, ANIK M. HARIATI3, ANDI KURNIAWAN3, DEWA G.R. WIADNYA3 1Doctoral Program, Faculty of Fisheries and Marine Science, Universitas Brawijaya. Jl. Veteran, Malang 65145, East Java, Indonesia. Tel.: +62-341-553512, Fax.: +62-341-557837, email: [email protected]/[email protected] 2Department of Aquaculture, Faculty of Fisheries, Agriculture and Biology, Universitas Bangka Belitung. Jl. Kampus Peradaban Balunijuk, Merawang Bangka 33172, Bangka Belitung Islands, Indonesia 3Faculty of Fisheries and Marine Science, Universitas Brawijaya. Jl. Veteran, Malang 65145, East Java, Indonesia. Tel.: +62-341-553512, Fax.: +62-341-557837, email: [email protected] Manuscript received: 30 November 2020. Revision accepted: 16 January 2021. Abstract. Kurniawan A, Hariati AM, Kurniawan A, Wiadnya DGR. 2021. First genetic record and the phylogenetic relationship of Osteochilus spilurus (Cyprinidae: Labeoninae) originating from Bangka and Belitung Islands, Indonesia. Biodiversitas 22: 794-802. Osteochilus spilurus is a freshwater fish from Southeast Asia. Species identification for this fish in Indonesia is still solely limited to morphological characteristics. Therefore it is highly important to validate using short DNA sequences that have been proven to be fast, effective, and accurate. Fish specimens were collected from Lenggang river in Belitung island, and Lebak river in Bangka island, Indonesia. Mitochondrial cytochrome b gene (cyt b, 408 bp) was amplified and sequenced. We collected fish specimens from Lenggang River in Belitung Island and Lebak River in Bangka Island, Indonesia. The results suggested that individuals from both islands are clustered together with a strong bootstrap value (100%) in a monophyletic clade. The specimens deposited at NCBI as the first Indonesian genetic records. They have a sister lineage to individuals from Sabah, Malaysia, although supported by a low bootstrap value. While geographical connection during the Pleistocene period allowed wide distribution of the species, differences in the evolutionary processes of historical biogeography and environmental changes are the predicted cause of the low genetic similarity. These findings can be used as a sequence reference and basic information for conservation of this species. Keywords: Cytochrome b, mtDNA, Osteochilus, Sundaland INTRODUCTION Fakhrurrozi et al. 2016; Kurniawan et al. 2020a). This fish is known by almost all Belitung people but is only known Osteochilus spilurus is native freshwater fish in by anglers on other islands. Belitung community likes to Indonesia, widely distributed in Southeast Asia. Previous consume this slightly bitter-tasting fish as a protein source studies have shown that geographic distribution of this (Kurniawan et al. 2016). Even these fish are included in species is Sundaland, including in Thailand, Malay society's culture with the emergence of ethnic songs and Peninsula, Borneo, and Sumatra Island (Kottelat et al. Batik with the theme of Cempedik. 1993; Fahmi et al. 2015; Haryono and Wahyudewantoro The economic value of O. spilurus has increased due to 2020; Lumbantobing and Huckstorf 2020). They are demand and habit of Belitung residents to consume it, usually found in acidic water of low-velocity rivers. The especially in East Belitung (Kurniawan and Triswiyana fish marked black blotch at the base of the tail and has a 2019). This fish, which was the least concern, now needs length of not more than 10 cm are benthopelagic (live serious attention due to the exploitation of nature and its between the surface and deep water areas). They also like trade. Consumption of fish relying on the natural catch can to move in groups with other small fish and are positioned pressure its natural population. Pragmatically, fish at the bottom of the group. There are no publications that domestication will reduce the burden on the natural specifically discuss O. spilurus apart from Bangka and ecosystem through fishing while satisfying consumer Belitung Islands. Most of the publications indicate the demand. Kurniawan et al. (2019) have started the first level existence of this species as an ichthyofauna in the domesticated as aquaculture commodities, and related freshwater river. aspects of fish domestication are studied in more detail by Osteochilus spilurus is not a common fish in Indonesia. Radona et al. (2020). At present, O. spilurus is in the first They have not got a national tag. For this species, each stage of domestication. Fish have been able to live in island has a local name. They are Toros in Malaysia, artificial containers on this level. Treatment of running Seluang Licin in Central Borneo, Seluang Sungai in water is a critical finding in the domestic process southern Sumatra, Kepait in Bangka island, and Cempedik (Kurniawan et al. 2019). Stress and fish mortality in Belitung island (Huckstorf 2012; Muslih et al. 2014; decreased when the water in the container flowed even KURNIAWAN et al. – First genetic record and phylogenetic relationship of Osteochilus spilurus 795 though in low debit. Malaysia, with 408 base pair (bp) cytochrome b (Cyt b) However, there is a gap problem that has not been gene primary (Esa et al. 2012). A fragment of the prepared before entering domestication. This species has mitochondrial cytochrome b (Cytb) gene is a frequently not been identified molecularly. Nowadays, the used marker for reconstructing species-level relationships. identification of O. spilurus in Indonesia is still limited to Cyt b has been used for species identification and its morphological characters (Kurniawan et al. 2020b; phylogenetic studies (Hsieh et al. 2001). In this study, we Mustikasari et al. 2018). The morphological approach has a used the 408 bp region of the mitochondrial cytochrome b weakness because a phenotype change in fish may cause an gene to equate the cytochrome b fragments used in the uncertain deviation in species validation. Although the previous record by Esa et al. (2012). This finding will be a meristic character is according to the description of Weber valuable benefit for species validation and future and de Beaufort (1916), O. spilurus from Bangka and identification references. The absence of O. spilurus gene Belitung island has a significant difference in eight records from Indonesia in the GenBank provides an morphometric characters and the body shape (Kurniawan et opportunity for this research to produce the first record. al. 2020b; Kurniawan et al. 2020c). The cause of the Gene bank records are expected to reflect increased morphological variation cannot be ascertained when the attention to O. spilurus, which is considered the least fish originated from two different islands. However, when concern in other Southeast Asia regions. Still, it is possible genetic variations cause phenotypic variations, it can refute at a more dangerous level to the population of Belitung or complement the argument for ecological adaptation. Its Island, especially East Belitung. molecular information needed in preparation for the breeding stage of domestication. Genetic records are useful for comparing subsequent identifications, considering MATERIALS AND METHODS genetic diversity, and compiling phylogeography across regions. Genetic identification is necessary to certainty and Sample source more specific species identification where traditional Live fish specimens were collected from the catches of methods cannot be applied (Ward et al. 2009). local fishermen in Kepulauan Bangka-Belitung Province, Mitochondrial DNA genes, which are widely used in Indonesia. The sample location was determined by interview fish identification, can also molecularly validate O. and verified before sampling from any single source. Random spilurus species. Bangka and Belitung, the two biggest samples of each wild Osteochilus spilurus were collected in islands in Bangka-Belitung Islands Province and have Lebak River, Payabenua Village, Merawang Sub-district, economic use in one of them, require molecular Bangka District, Bangka Island in March 2019 (2°04'21.4"S, investigation to confirm their phylogenetic position. 105°52'12.0"E) and in Lenggang River, Lintang Village, However, the lack of genetic studies of this species makes Simpang Renggiang Sub-district, East Belitung District, it necessary to adjust the type of gene used in molecular Belitung Island in May 2019 (2°55'09.5"S 108°06'35.4"E) identification. There is only one database source of O. (Figure 1). The species did not comprise stock requiring spilurus genetic sequences available at the National Center special permits because it is included in the least concerned for Biotechnology Information (NCBI 2019). The category in IUCN (Huckstorf 2012). The fishing gear used nucleotides are a specimen of Tawau River, Sabah, adjusts to the habits of local fishers. Figure 1. Sampling location of Osteochilus spilurus showing at Bangka and Belitung Islands, Indonesia 796 BIODIVERSITAS 22 (2): 794-802, February 2021 Procedures rpm at cold temperature for 10 minutes. Three hundred Sample preservation (300) µl of chloroform/isoamyl alcohol (24:1) was added to Thirty (30) individuals were collected from Bangka and 100 µl of the supernatant, and the result was centrifuged for
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