Systematics of the subfamily Danioninae (Teleostei: Cypriniformes: Cyprinidae).

The members of the cyprinid subfamily Danioninae form a diverse and scientifically important group of fishes, which includes the zebrafish, Danio rerio. The diversity of this assemblage has attracted much scientific interest but its monophyly and the relationships among its members are poorly understood. The phylogenetic relationships of the Danioninae are examined herein using sequence data from mitochondrial cytochrome b, mitochondrial cytochrome c oxidase I, nuclear opsin, and nuclear recombination activating gene 1. A combined data matrix of 4117 bp for 270 taxa was compiled and analyzed. The resulting topology supports some conclusions drawn by recent studies on the group and certain portions of the traditional classification, but our results also contradict key aspects of the traditional classification. The subfamily Danioninae is not monophyletic, with putative members scattered throughout Cyprinidae. Therefore, we restrict Danioninae to the monophyletic group that includes the following genera: Amblypharyngodon, Barilius, Cabdio, Chela, Chelaethiops, Danio, Danionella, Devario (including Inlecypris), Esomus, Horadandia, Laubuca, Leptocypris, Luciosoma, Malayochela, Microdevario, Microrasbora, Nematabramis, Neobola, Opsaridium, Opsarius, Paedocypris, Pectenocypris, Raiamas, Rasbora (including Boraras and Trigonostigma), Rasboroides, Salmostoma, Securicula, and Sundadanio. This Danioninae sensu stricto is divided into three major lineages, the tribes Chedrini, Danionini, and Rasborini, where Chedrini is sister to a Danionini-Rasborini clade. Each of these tribes is monophyletic, following the restriction of Danioninae. The tribe Chedrini includes a clade of exclusively African species and contains several genera of uncertain monophyly (Opsarius, Raiamas, Salmostoma). Within the tribe Rasborini, the species-rich genus Rasbora is rendered non-monophyletic by the placement of two monophyletic genera, Boraras and Trigonostigma, hence we synonymize those two genera with Rasbora. In the tribe Danionini, the miniature genus Danionella is recovered as the sister group of Danio, with D. nigrofasciatus sister to D. rerio.

Annotated checklist of freshwater fishes from Brunei Darussalam, Borneo.

The freshwater fish diversity of Brunei Darussalam is reviewed. At present, a total of 104 species of freshwater fishes from 59 genera, 24 families and 10 orders is recorded. Eight species including one introduced species and two from brackish waters present new records for Brunei. The family Cyprinidae has the highest diversity with 40 species. Forty-six species are believed to be endemic to Borneo including six species known only from Brunei Darussalam namely Barbodes xouthos, Rasbora tubbi, Gastromyzon cranbrookii, Gastromyzon venustus, Neogastromyzon brunei and Ompok platyrhynchus and three are exotics.

Eco-hydromorphic Classification for Understanding Stream Macroinvertebrate Biodiversity in Brunei Darussalam, Northern Borneo.

Kate Baker, Michael A. Chadwick, and Zohrah Haji Sulaiman (2016) Linking ecology with river geomor- phology and hydrology (geomorphic and hydraulic template) plays an important role in the study of macroinver- tebrate biodiversity. This understanding and knowledge is crucial in implementing sensible conservation management for ecosystem health monitoring. However, most macroinvertebrate research has been conducted in temperate ecosystems. This study examines the eco-hydrogeomorphology and macroinvertebrate biodiversity of two remote tropical streams in northern Borneo (Bukit Pagon catchment, Brunei Darussalam's highest mountain - 1850 m) using temperate classification models, more specifically, biotopes. Fast flowing biotopes were defined as bedrock runs and cobble riffles whilst the slow flowing biotopes were deposition pools. Macroinvertebrate size structure associated with biotopes, which can influence overall ecological processes, was also investigated. Forty-three macroinvertebrate taxa were recorded during the study; biodiversity was similar between the study streams. There were differences among biotopes with the lowest diversity occurring in fast flowing biotopes (p = 0.05*). Community structure also varied among the biotopes. Cluster analysis of macroinvertebrate abundance revealed an 0.8 dissimilarity between the fast and slow biotopes. Several taxa were found in multiple biotopes, which is likely linked to the occurrence of moss and leaf litter. Macroinvertebrate size structure distribution between the fast and slow biotopes was statistically different. Our findings suggest biotopes may be an appropriate scale to investigate macroinvertebrate biodiversity in tropical streams. Specifically, we found that biotopes had different macroinvertebrate communities and richness. Further research is required to understand the importance of habitat parameters that are not directly related to flow velocities such as moss. These habitats are important as places of refuge, allowing colonisation that would otherwise be inhospitable during flood periods.

Genetic identification and structure of Clarias batrachus (Linnaeus, 1758) from Southeast Asia using a mitochondrial DNA marker.

A phylogenetic tree and median-joining network based on cytochrome b sequence data revealed clades consistent with morphological differences and geographical distribution of Clarias batrachus (Linnaeus, 1758) in Southeast Asia. AMOVA analysis for variation was significant among populations (P<0.05) and was in agreement with morphological differences. Pairwise differences were significant between Java and Brunei/Borneo, Brunei/Borneo and west Malaysia, and Java and west Malaysia samples (P < 0.05). Closest relationships were found between samples from Brunei/Borneo and Java, and between west Malaysia and Laos-Sumatra. Nine haplotypes were unique to geographical regions. The Java species had high haplotype (1.000 ± 0.126) but low nucleotide (0.017) diversities, suggesting a population bottleneck followed by expansion. However, SSD and Hri (P=0.5) did not support demographic expansion. Instead, purifying selection where mutations occur and accumulate at silent sites is a more acceptable explanation.

Molecular analysis of bacterial diversity in mudflats along the salinity gradient of an acidified tropical Bornean estuary (South East Asia).

BACKGROUND: The Brunei River and Bay estuarine system (BES) in the northwest of Borneo is acidic and highly turbid. The system supports extensive intertidal mudflats and presents a potentially steep salinity and pH gradient along its length (45 km). Temporal variation in physical parameters is observed diurnally due to seawater flux during tidal forcing, and stochastically due to elevated freshwater inflow after rains, resulting in a salinity range between 0 and 34 psu. High velocity freshwater run-off from acid sulphate formations during monsoon seasons results in highly variable and acidic conditions (pH 4) at the upper reaches of the BES, whereas the pH is relatively stable (pH 8) at the seaward extremes, due to mixing with seawater from the South China Sea. At their surfaces, the BES mudflats present microbial ecosystems driven by oxygenic phototrophs. To study the effect of various physical parameters on the bacterial diversity of the BES mudflats, surface samples were collected from six sites stretching over 40 km for molecular and phylogentic analysis. RESULTS: The bacterial diversity at these sites was compared by community fingerprinting analysis using 16S rRNA gene based denaturing gradient gel electrophoresis and by 16S rRNA gene sequencing and phylogenetic analyses. Results revealed functionally conserved, diatom-driven microbial mudflat communities composed of mainly novel, uncultured species. Species composition was evaluated as 50-70% unique for each site along the BES. Clustering of the sequences commonly occurred and revealed that proteobacterial diversity was related to the salinity gradient. When considering all phyla, the diversity varied consistently with physical parameters (including anthropogenic) that are expected to influence microbial composition. CONCLUSION: The BES mudflats were found to comprise the typical functional groups of microorganisms associated with photosynthetic carbon flux, sulfur cycling (Gamma- and Deltaproteobacteria), and decomposition (Bacteroidetes). From a structural perspective, however, the mudflats constituted discretely distributed communities along the physical gradient of the BES, composed of largely novel species of Bacteria. This study provides first insights into patterns of bacterial community structure in tropical South East Asian coastal ecosystems that are potentially threatened by increasing variability in pH and salinity, in line with predicted future environmental change.

Genetic population structure of red snapper (Lutjanus malabaricus) and orange-spotted grouper (Epinephelus coiodes) in Brunei and Sabah.

The genetic population structure of red snapper Lutjanus malabaricus and orange-spotted grouper Epinephelus coiodes in Brunei and Sabah was investigated using allozyme electrophoresis. Samples were collected from three sites in Brunei for E. coiodes and from three sites in Brunei and Sabah for L. malabaricus. A total of 22 loci and 16 loci were scored, respectively. The index of fixation (F(ST) ) for the E. coiodes population was 0.176 but, in general, it lacked within-population structuring. The F(ST) was particularly high between Brunei Bay/Jerudong and Brunei Bay/ Kuala Belait, suggesting genetic subdivision on a small spatial scale. Isolation of Brunei Bay from the South China Sea may constrain the movement of adult fishes and larval dispersal, thereby reducing homogeneity among subpopulations. All variable loci for E. coiodes were in Hardy-Weinberg equilibrium except for MDH* and GTDHP* (P < 0.01), in which two subpopulations showed an excess of heterozygotes (P < 0.01). The study on the L. malabaricus population showed a heterozygote deficit of approximately 60% in variable loci (F(ST) genetic variation within population = 0.45; P < 0.05); however, the mean observed heterozygosity for the population far exceeded L. malabaricus populations in Australia and Indonesia. A F(ST) value of 0.076 revealed moderate genetic differentiation among subpopulations of L. malabaricus. The genotypes were likely to be drawn from the same distribution in Jerudong and Kuala Belait. This study infers that sustainable management of snapper and grouper resources in Brunei waters must take into account the presence of a single stock and two stocks, respectively.

Phylogenetic relationships of Danio within the order Cypriniformes: a framework for comparative and evolutionary studies of a model species.

The evolutionary relationships of species of Danio and the monophyly and phylogenetic placement of the genus within the family Cyprinidae and subfamily Rasborinae provide fundamentally important phyloinformatics necessary for direct evaluations of an array of pertinent questions in modern comparative biology. Although the genus Danio is not one of the most diverse within the family, Danio rerio is one of the most important model species in biology. Many investigations have used this species or presumed close relatives to address specific questions that have lasting impact on the hypothesis and theory of development in vertebrates. Largely lacking from this approach has been a holistic picture of the exact phylogenetic or evolutionary relationships of this species and its close relatives. One thing that has been learned over the previous century is that many organismal attributes (e.g., developmental pathways, ecologies, behaviors, speciation) are historically constrained and their origins and functions are best explained via a phylogenetic approach. Herein, we provide a molecular evaluation of the phylogenetic placement of the model species Danio rerio within the genus Danio and among hypothesized closely related species and genera. Our analysis is derived from data using two nuclear genes (RAG1, rhodopsin) and five mitochondrial genes (ND4, ND4L, ND5, COI, cyt b) evaluated using parsimony, maximum likelihood, and Bayesian analyses. The family Cyprinidae is resolved as monophyletic but the subfamily Rasborinae (priority over Danioinae) is an unnatural assemblage. Danio is identified as a monophyletic group sister to a clade inclusive of the genera Chela, Microrasbora, Devario, and Inlecypris, not Devario nor Esomus as hypothesized in previous studies. Danio rerio is sister to D. kyathit among the species of Danio evaluated in this analysis. Microrasbora and Rasbora are non-monophyletic assemblages; however, Boraras is monophyletic.

Mitochondrial DNA sequence analyses in Bornean sucker fishes (Balitoridae: Teleostei: Gastromyzontinae).

Phylogenetic relationships among Bornean sucker fishes (Teleostei: Balitoridae: Gastromyzontinae) were investigated by comparing cytochrome b gene sequences from eight species. The results were in general agreement with previous morphology-based studies. It was found that the genera Gastromyzon and Neogastromyzon are both monophyletic and that the Chinese homalopterid Crossostoma lacustre (Homalopterinae) is not related to the Bornean species. This molecular-level study of cytochrome b gene variation in Bornean gastromyzontins will undoubtedly help to shed light on the molecular systematics of this unique freshwater fish.