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Benthic species, though ecologically important, are vulnerable to genetic loss and population size reduction due to impacts from fishing trawls. An assessment of genetic diversity and population structure is therefore needed to assist in a resource management program. To address this issue, the two-spined yellowtail stargazer (Uranoscopus cognatus) was collected within selected locations in the Indo-West Pacific (IWP). The partial mitochondrial DNA cytochrome c oxidase subunit 1 and the nuclear DNA recombination activating gene 1 were sequenced. Genetic diversity analyses revealed that the populations were moderately to highly diversified (haplotype diversity, H = 0.490–0.900, nucleotide diversity, π = 0.0010–0.0034) except sampling station (ST) 1 and 14. The low diversity level, however was apparent only in the matrilineal marker (H = 0.118–0.216; π = 0.0004–0.0008), possibly due to stochastic factors or anthropogenic stressors. Population structure analyses revealed a retention of ancestral polymorphism that was likely due to incomplete lineage sorting in U. cognatus, and prolonged vicariance by the Indo-Pacific Barrier has partitioned them into separate stock units. Population segregation was also shown by the phenotypic divergence in allopatric populations, regarding the premaxillary protrusion, which is possibly associated with the mechanism for upper jaw movement in biomechanical feeding approaches. The moderate genetic diversity estimated for each region, in addition to past population expansion events, indicated that U. cognatus within the IWP was still healthy and abundant (except in ST1 and 14), and two stock units were identified to be subjected to a specific resource management program.

The taxonomic status of the Southeast Asian spotted barb, Barbodes binotatus (Teleostei: Cyprinidae), has puzzled researchers because of large but inconsistent geographic variation of its body melanin marking pattern. In this study, the authors appraise the differentiation of B. binotatus and two closely related species, Barbodes rhombeus and saddle barb, Barbodes banksi, in Peninsular Malaysia using mitochondrial and nuclear markers. The results of this study reveal that the Peninsular Malaysia populations of each of the three species form largely reciprocal monophyletic lineages that differ from each other by a minimum of 2.3% p-genetic distance using COI gene. Nonetheless, specimens of B. binotatus in Peninsular Malaysia are only distantly related to specimens of B. binotatus in Java (type locality). The monophyly of B. banksi is not refuted although specimens of Peninsular Malaysia are genetically distinct from those of Sarawak (type locality). The authors discuss alternative hypotheses whether each of these three valid species is a single species or each of the main five genetic lineages revealed in this study represents a distinct species. Preliminary investigations reveal a mito-nuclear discordance at one locality in Peninsular Malaysia where B. binotatus and B. banksi co-occur. Further studies should inform on the extent of reproductive porousness between these two lineages and others.

The tire track eel, Mastacembelus favus, is an economically important freshwater fish in mainland Southeast Asia. Understanding the spatial population structure is important to design an effective and efficient management of the species. The aim of this study was to investigate the genetic diversity and structure of M. favus populations. In total, 519 individuals from 30 locations across its distribution range were genotyped using a set of eight polymorphic microsatellite loci. Results showed that populations in Mekong and Chao Phraya exhibited higher genetic variability than those from Gulf of Thailand and Malay–Thai Peninsula. Clustering results revealed two evolutionarily isolated groups geographically separated at Northeastern Cambodia; the more widespread group was further divided into two subclusters. The spatial genetic variation and structure of its populations are attributed to the evolutionary history and population dynamics of the species. The study provides important information for the management and conservation of M. favus in Southeast Asia.

Genus Rusa, belonging to the deer family Cervidae is native to the Indo-Malaya Archipelago (IMA). However, detailed information on the Rusa genus in the IMA is limited. This review provides comprehensive information on the Rusa genus in the IMA including, threats and conservation efforts. There are four species of deer in Rusa genus, which is Sambar deer (Rusa unicolor), Javan deer (Rusa timorensis), Visayan spotted deer (Rusa alfredi) and Philippine deer (Rusa marianna). Despite their wide distribution in the South Asian and Southeast Asian regions, they are under serious threats. Some conservation efforts that are being done to protect and conserve them among others are; 1) facilities protection, 2) habitat enrichment programme, 3) Ex-situ conservation, 4) legislations, and 5) captive breeding. Conservation through genetics is also an important step in conserving these species. Recommendations for conservation of the genus are also discussed; 1. maintenance of ecosystem. 2. more effective monitoring system on the existing protected area. 3. ex-situ conservation, and 4. habitat monitoring.

The Japanese scad Decapterus maruadsi (Carangidae) is an economically important marine species in Asia but its exploitation shows signs of overfishing. To document its stock structure, a population...

The fish genus Notopterus Lacepède, 1800 (Notopteridae) currently includes only one species, the Asian bronze featherback Notopterus notopterus (Pallas, 1769). This common freshwater species is widely distributed in the Oriental region, from the Indus basin in the west, the Mekong basin in the east and Java Island in the south. To examine the phylogeographic structure of N. notopterus across its range, we analysed 74 publicly available cytochrome oxidase I (COI) sequences, 72 of them determined from known-origin specimens, along with four newly-determined sequences from Peninsular Malaysian specimens. We found that N. notopterus is a complex of two allopatric species that diverge from each other by 7.5% mean p-distance. The first species is endemic to South Asia (from Indus basin to Ganga-Brahmaputra system), whereas the distribution of the second species is restricted to Southeast Asia. The exact limit between the distributions of these two species is not known, but it should fall somewhere between the Ganga-Brahmaputra and Salween basins, a region already identified as a major faunal boundary in the Oriental region. The name N. notopterus is retained for the Southeast Asian species, while the name Notopterus synurus (Bloch & Schneider, 1801) should be applied to the South Asian species. A comparative morphological study is needed to reveal the degree of morphological differentiation between the two species.

This study utilized genetic and morphometric approaches to assess the molecular and morphometric differentiation among commercially important species of mud crab. Molecular investigations were derived from 542 bp mitochondrial DNA COI on 249 individuals within genus Scylla from nine states in Malaysia represents four marine regions; South China Sea, Sulu Sea, Straits of Singapore and Straits of Malacca. Four specimens were obtained from Indonesia to give a robust analysis in this study. For species delimitation, Automatic Barcode Gap Discovery (ABGD) method on a web interface was employed. Analysis on phylogenetics was implemented utilizing Neighbour joining (NJ) and Maximum Parsimony (MP) methods. The inter- and intraspecies genetic distances (Ds) was computed using Kimura 2-parameter distance and executed in MEGA version 5.05. All samples were genetically and morphologically identified and clustered into four distinct species. Among the species, S. olivacea was the most abundant (n = 111), on the other hand the occurrence of S. paramamosain in Malaysia was very low (n = 29). No single individual of S. serrata from Malaysia was recorded in this study. Both genetic distance and phylogenetic approaches exhibited a correlative monophyletic association among all specimens analysed. This present study is crucial as it reports the reassessment of all species within genus Scylla in Malaysia, eventually could be employed as a reference source for subsequent research mainly on mariculture and other conservation efforts for the species. Keywords: Scyla spp., Mud crab, Morphological identification, Molecular identification, COI

The first molecular phylogeny for mountain reed snakes (genus Macrocalamus) based on the mitochondrial gene cytochrome b is not entirely consistent with the previous taxonomy based on morphology and colour pattern. Macrocalamus chanardi is shown to be a species complex composed of three different allopatric lineages distributed across different upland areas in Peninsular Malaysia that are morphologically conserved but genetically distinct. A new and morphologically different species, Macrocalamus emas sp. nov., is described from the Cameron Highlands, Pahang, Peninsular Malaysia. It occurs in sympatry with four other ecologically equivalent species of Macrocalamus and one other species of Collorhabdium. The phylogeographical pattern of sympatric genetically distinct species of Macrocalamus endemic to upland areas is attributed to the fossorial nature of these snakes and the montane forest expansion and retraction resulting from cyclical, glacioeustatically driven climatic processes that have reconstructed the geography of Sundaland continuously over the last 25 Myr.

This study was carried out to evaluate the effect of mannan oligosacharide (MOS) as dietary supplement on haematological parameters, immune response and protection against Aeromonas hydrophila in striped catfish (Pangasianodon hypophthalmus, Sauvage, 1878) juveniles. Triplicate groups of juvenile striped catfish (initial weight 20.41 ± 1.64 g) were fed twice per day at 2.5% of body weight for 12 weeks, with 0 (control), 0.2, 0.4, 0.6 or 0.8% MOS diets. At the end of the feeding trial, fish were intraperitoneally challenged with 1 × 106 CFU ml−1of A. hydrophila and mortalities were recorded over 21 days. The haematological and immunological parameters were assessed both in pre- and post-challenged fish groups. Dietary inclusion of MOS did not show any significant influence on the majority of the haematological parameters of striped catfish when they were not infected. But it showed a significant positive influence on some of the most important parameters, namely RBC, WBC, lymphocyte, granulocyte and lysozyme activity and survival, after a 2-week challenge with A. hydrophila. After 2-week infection with A. hydrophila, a better concentration of red blood cell (RBC), white blood cell (WBC), granulocyte, total Ig content, lysozyme activity and survival were observed when striped catfish were fed with MOS at 0.4% and above, with significantly highest at 0.6% MOS diet than the control diet. Based on the results of this study, it is concluded that supplementation of MOS at least 0.4% is sufficient to activate the immune response and makes the striped catfish more resistant to A. hydrophila infection.

The complex climatic and geological history of Southeast Asia has been hypothesised to determine the most important aspects of the current phylogeographical structure and distribution of living organisms throughout the region. To test existing hypotheses, the genetic structure of the tire track eel, Mastacembelus favus, was investigated using 823 bp of mitochondrial DNA cytochrome b from 469 individuals from 51 localities encompassing its native range. The results classified all haplotypes into two major lineages, Lineage 1, which was further divided into Lineages 1a (lower Mekong, eastern Gulf of Thailand and Malay—Thai Peninsula), 1b (Banpakong River), 1c (Chao Phraya, Gulf of Thailand and Malay—Thai Peninsula) and 1d (Khlang Yai River), and Lineage 2, the upper reaches of the lower Mekong and the middle Mekong. Strong genetic discontinuities dated approximately 5 MYA were discovered in the Mekong with limited geographical overlap, suggesting a historically dissected drainage between two sections and species colonisation via different routes. The widespread Lineage 1 showed a strong signature of population expansion during the Pleistocene climate oscillation. Haplotype characteristics in the Malay—Thai Peninsula are hypothesised to result from postglacial dispersal from the Mekong and Chao Phraya through an extended Pleistocene drainage network.

A total of 74 shrimp specimens were sequenced at a 584 bp segment of the cytochrome oxidase subunit I (COI) gene to examine patterns of DNA barcode variation in a mangrove biodiversity hotspot. The Maximum Likelihood tree, barcode gap analysis, Automatic Barcode Gap Discovery analysis and sequence comparisons with data available from Barcode of Life Data System and GenBank recovered 18 taxa of which 15 were identified to species level, 2 at genus level and a single taxon at order level. Two deep mitochondrial DNA lineage divergences were found in the giant tiger prawn

The population genetic diversity and demographic history of the longtail tunaThunnus tonggolin Malaysian waters was investigated using mitochondrial DNA D-loop and NADH dehydrogenase subunit 5 (ND5). A total of 203 (D-loop) and 208 (ND5) individuals ofT. tonggolwere sampled from 11 localities around the Malaysian coastal waters. Low genetic differentiation between populations was found, possibly due to the past demographic history, dispersal potential during egg and larval stages, seasonal migration in adults, and lack of geographical barriers. The gene trees, constructed based on the maximum likelihood method, revealed a single panmictic population with unsupported internal clades, indicating an absence of structure among the populations studied. Analysis on population pairwise comparison ФSTsuggested the absence of limited gene flow among study sites. Taken all together, high haplotype diversity (D-loop = 0.989–1.000; ND5 = 0.848–0.965), coupled with a low level of nucleotide diversity (D-loop = 0.019–0.025; ND5 = 0.0017–0.003), “star-like” haplotype network, and unimodal mismatch distribution, suggests a recent population expansion for populations ofT. tonggolin Malaysia. Furthermore, neutrality and goodness of fit tests supported the signature of a relatively recent population expansion during the Pleistocene epoch. To provide additional insight into the phylogeographic pattern of the species within the Indo-Pacific Ocean, we included haplotypes from GenBank and a few samples from Taiwan. Preliminary analyses suggest a more complex genetic demarcation of the species than an explicit Indian Ocean versus Pacific Ocean delineation.

Knowledge on the precise identification of fish resources is critical for sustainable fisheries management. This study employs the DNA barcoding approach to generate a molecular taxonomic catalogue of commercially important reef fishes in the waters of Weh Island (Aceh Province), the most northerly inhabited island in the biodiverse Indonesian Archipelago. The waters not only support artisanal fisheries but also a feeder for the industry in the greater island of Aceh. In total, 230 specimens from 72 species belonging to 32 genera and 17 families were DNA barcoded, representing a major segment of the captured reef fish taxa and a quarter of fish species diversity that had previously been recorded. The sequence read lengths were 639 bp revealing 359 conserved sites, 280 variable sites, 269 parsimony informative and 11 singletons. Our molecular findings paralleled the morphological identification with no evidence of cryptic species or new species discovery. This study is a significant contribution to the fisheries statistics of this area, which would facilitate assessment of species catch composition and hence for strategizing management plans. It is an important input to the DNA barcode library of Indonesian marine fishes and to the global DNA barcode entries in general.

The tropical freshwater bighead catfish Clarias macrocephalus is widely distributed in Southeast Asia and is economically important in both the aquaculture and fisheries industries. In the current study, eight polymorphic microsatellite markers were employed to evaluate the genetic variation of 15 C. macrocephalus populations from Peninsular Malaysia, and the Mekong Delta populations of Cambodia and Vietnam. Overall, the number of alleles in C. macrocephalus populations ranged from 1 to 25 alleles. In general, genetic diversity was higher in the mainland populations (Cambodia, HE = 0.761 to 0.813 and Vietnam, HE = 0.759 to 0.789 with mean number of alleles of 10.13–13.88) compared to Peninsular Malaysia (HE = 0.519 to 0.699 with mean number of alleles of 4.5–10.75). High linkage disequilibrium and deviation from HWE suggest inbreeding in most Peninsular Malaysia populations which was also reflected by significant FIS values (0.217 to 0.410, p

Aquatic ecosystems that form major biodiversity hotspots are critically threatened due to environmental and anthropogenic stressors. We believe that, in this genomic era, computational methods can be applied to promote aquatic biodiversity conservation by addressing questions related to the evolutionary history of aquatic organisms at the molecular level. However, huge amounts of genomics data generated can only be discerned through the use of bioinformatics. Here, we examine the applications of next-generation sequencing technologies and bioinformatics tools to study the molecular evolution of aquatic animals and discuss the current challenges and future perspectives of using bioinformatics toward aquatic animal conservation efforts.

Data are available on Dryad with the DOI 10.5061/dryad.5j501., The complex climatic and geological history of Southeast Asia has shaped this region’s high biodiversity. In particular, sea level fluctuations associated with repeated glacial cycles during the Pleistocene both facilitated, and limited, connectivity between populations. In this study, we used data from two mitochondrial and three anonymous nuclear markers to determine whether a fresh/brackish water killifish, Aplocheilus panchax, Hamilton, 1822, could be used to further understand how climatic oscillations and associated sea level fluctuations have shaped the distribution of biota within this region, and whether such patterns show evidence of isolation within palaeodrainage basins. Our analyses revealed three major mitochondrial clades within A. panchax. The basal divergence of A. panchax mitochondrial lineages was approximately 3.5 Ma, whilst the subsequent divergence timings of these clades occurred early Pleistocene (~2.6 Ma), proceeding through the Pleistocene. Continuous phylogeographic analysis showed a clear west-east dispersal followed by rapid radiation across Southeast Asia. Individuals from Krabi, just north of the Isthmus of Kra, were more closely related to the Indian lineages, providing further evidence for a freshwater faunal disjunction at the Isthmus of Kra biogeographic barrier. Our results suggest that Sulawesi, across the Wallace Line, was colonised relatively recently (~30 ka). Nuclear DNA is less geographically structured, although Mantel tests indicated that nuclear genetic distances were correlated with geographic proximity. Overall, these results imply that recent gene flow, as opposed to historical isolation, has been the key factor determining patterns of nuclear genetic variation in A. panchax, however, some evidence of historical isolation is retained within the mitochondrial genome. Our study further validates the existence of a major biogeographic boundary at the Kra Isthmus, and also demonstrates the use of widely distributed fresh/brackishwater species in phylogeographic studies, and their ability to disperse across major marine barriers in relatively recent time periods., Global Innovation Initiative Grant to MdB, "Environmental DNA for rapid assessment of biodiversity and conservation priorities in Indonesia", will be used for data submission in Dryad upon acceptance of publication

Zamroni Y, Soewardi K, Suryobroto B, Jaafar Z. 2016. Short Communication: Conservation of mangrove gobies in LesserSunda Islands, Indonesia. Biodiversitas 17: 553-557. Ecosystems goods and services from mangrove forests are especially vital to coastal communities. Yet mangrove areas continue to be deforested at unprecedented rates. Using gobioid fishes associated with mangrove forests as focal organisms, we assessed their diversity in 14 selected sites within the Lesser Sunda group of islands. We applied Correspondence analysis to determine the relationships between ecosystems based on the occurrence of these fishes and complementarity analysis to identify the minimum number of sites to conserve maximum diversity based on a rarity algorithm. We recovered 55 gobioid fish species at these mangrove areas, and proposed six mangrove areas within the Lesser Sunda group of islands as areas of conservation priority: Loh Sebita, Oebelo, Bipolo, Lembar Bay, Selindungan, and Kawangu. The three former areas are already within protected zones while the remaining latter three areas are at present unprotected. The argument for the conservation of these threeremaining areas is a compelling one, based on our data (diversity of gobioid fishes), and corroborating data (diversity of corals, reef fishes, stomatopods, seagrasses, and marine birds) from other studies.Keywords: Brackish, mangroves, deforestation, Nusa Tenggara, Threatened Species, Gobiidae

DNA barcoding of the cytochrome oxidase subunit I (COI) gene was utilized to assess the species diversity of the freshwater halfbeak genus Hemirhamphodon. A total of 201 individuals from 46 locations in Peninsular Malaysia, north Borneo (Sarawak) and Sumatra were successfully amplified for 616 base pairs of the COI gene revealing 231 variable and 213 parsimony informative sites. COI gene trees showed that most recognized species form monophyletic clades with high bootstrap support. Pairwise within species comparisons exhibited a wide range of intraspecific diversity from 0.0% to 14.8%, suggesting presence of cryptic diversity. This finding was further supported by barcode gap analysis, ABGD and the constructed COI gene trees. In particular, H. pogonognathus from Kelantan (northeast Peninsular Malaysia) diverged from the other H. pogonognathus groups with distances ranging from 7.8 to 11.8%, exceeding the nearest neighbor taxon. High intraspecific diversity was also observed in H. byssus and H. kuekanthali, but of a lower magnitude. This study also provides insights into endemism and phylogeographic structuring, and limited support for the Paleo-drainage divergence hypothesis as a driver of speciation in the genus Hemirhamphodon.

The family Sciaenidae, known as croakers or drums, is one of the largest perciform fish families. A recent multi-gene based study investigating the phylogeny and biogeography of global sciaenids revealed that the origin and early diversification of this family occurred in tropical America during the Late Oligocene-Early Miocene before undergoing range expansions to other seas including the Indo-West Pacific, where high species richness is observed. Despite this clarification of the overall evolutionary history of the family, knowledge of the taxonomy and phylogeny of sciaenid genera endemic to the Indo-West Pacific is still limited due to lack of a thorough survey of all taxa. In this study, we used DNA-based approaches to investigate the evolutionary relationships, to explore the species diversity, and to elucidate the taxonomic status of sciaenid species/genera within the Indo-West Pacific clade. Three datasets were herein built for the above objectives: the combined dataset (248 samples from 45 currently recognized species) from one nuclear gene (RAG1) and one mitochondrial gene (COI); the dataset with only RAG1 gene sequences (245 samples from 44 currently recognized species); and the dataset with only COI gene sequences (308 samples from 51 currently recognized species). The latter was primarily used for our biodiversity exploration with two different species delimitation methods (Automatic Barcode Gap Discovery, ABGD and Generalized Mixed Yule Coalescent, GMYC). The results were further evaluated with help of four supplementary criteria for species delimitation (genetic similarity, monophyly inferred from individual gene and combined data trees, geographic distribution, and morphology). Our final results confirmed the validity of 32 currently recognized species and identified several potential new species waiting for formal descriptions. We also reexamined the taxonomic status of the genera, Larimichthys, Nibea, Protonibea and Megalonibea, and suggested a revision of Nibea and proposed a new genus Pseudolarimichthys.