The U.S. National Science Foundation-funded (DEB 1022720) 'All Cypriniformes Species Inventory' was initiated in 2010 and will be completed in 2015. It has accelerated the rate of discovery and description of cypriniform fishes, expanded our knowledge of the phylogenetic relationships of cypriniforms, increased the capacity for systematic research in other countries through student training and establishing long-term collaborations, including conferences in Thailand in 2012, Brunei in 2013, Burundi in 2013, and Malaysia in 2014, led to the formation of the Asian Society of Ichthyologists, and made available large numbers of specimens and tissues of freshwater fishes, including many species never before collected, in permanent collections in foreign and U.S. institutions.
This is the first study to identify and determine the phylogenetics of neritids found in Malaysia. In total, twelve species from the family Neritidae were recorded. Ten species were from the genus Nerita and two species were from the genus Neritina. DNA barcodes were successfully assigned to each species. Although some of these species were previously reported in the region, three are only presently reported in this study. The dendrogram showed Nerita and Neritina strongly supported in their respective monophyletic clades. Phylogenetic positions of some species appeared unstable in the trees. This could be due to the differences in a small number of nucleotides, thus minimizing genetic variation between each specimen and species.
We evaluated genetic differentiation among ten presumed Japanese threadfin bream, Nemipterus japonicus populations along the coast of Peninsular Malaysia based on the partial sequence of the mitochondrial cytochrome b gene (982 bp). Genetic divergences (Kimura-2 parameter) ranged from 0.5% to 0.8% among nine of the ten populations while these nine populations were 4.4% to 4.6% diverged from the Kuala Besar population located at the Northeast coast. The constructed Neighbour Joining (NJ) phylogenetic trees based on haplotypes showed the Kuala Besar population forming an isolated cluster. The Analysis of Molecular Variance (AMOVA) of the ten populations a priori assigned into four regions, revealed that most of the variation occurred within population with a fairly low but significant level of regional differentiation (FST = 0.07, p < 0.05, FSC = 0.00, p > 0.05 and FCT = 0.07, p < 0.05) attributed to the Kuala Besar population. p Value after Bonferroni correction revealed that only pairwise FST values involving the Kuala Besar population with the other nine populations were significant. Thus, this study revealed that the N. japonicus populations off Peninsular Malaysia were panmictic. However, the Kuala Besar population, although morphologically identical was composed of a genetically discrete taxon from the rest. These findings are important contributions in formulating sustainable fishery management policies for this important fishery in Peninsular Malaysia.
A primary factor in population management and wildlife conservation is the delineation of population units derived from descriptions of population genetic structure. Yet, predicting factors that influence the patterns of gene flow in a population particularly at landscape scales remains a major challenge in evolutionary biology. Here we report a population genetic study of the mud crab Scylla olivacea examined based on a 542 bp segment of the mitochondrial DNA cytochrome c oxidase I gene among 91 individuals from six localities in the west and east coast of Peninsular Malaysia. In total 55 unique haplotypes were distinguished with 45 private haplotypes and a single common haplotype shared among all populations studied. The other ten haplotypes were shared among various populations. The sharing of this haplotype reflects the connection of the mangrove areas between east and west coast of Peninsular Malaysia. High haplotype diversity (h = 0.968 ± 0.021; mean ± SD) and low nucleotide diversity (π = 0.120 ± 0.015; mean ± SD) were displayed, which may be indicative of genetic bottleneck events. No significant phylogenetic lineages were recognized using neighbour-joining and maximum parsimony methods. Hierarchical AMOVA analysis indicated that 99.33 % of the genetic variation was contained within populations and 0.67 % occurred among populations, suggesting no geographical patterning among populations studied, supported by F st test. Mismatch distribution analysis showed that the observed distribution of the pairwise mutation differences among haplotypes was multimodal, which is not concordant with a sudden range expansion scenario. However, neutrality tests showed non-significant negative values suggesting that the populations studied may have experienced past population growth, but the expansion may have been restricted to separate local areas that resulted in the non-significant negative Fu's Fs and Tajima's D value. Overall, this present preliminary study was able to be a reference on the phylogenetic relationships and assessment of genetic structure of Scylla sp. in Malaysia.
The white-bellied sea eagle, Haliaeetus leucogaster, displays reversed sexual size dimorphism and is monomorphic for adult plumage coloration. Early attempts to identify sex in sexually monomorphic birds were based on morphological or chromosomal characters, but since avian W-specific DNA sequences were identified, PCR amplification has become commonly used for molecular sexing. We used a PCR test employing primers that amplify two homologous fragments of both the CHD-W gene, unique to females, and the CHD-Z gene, occurring in both sexes. This test was applied to five individuals of H. leucogaster from the Malacca Zoo and to male and female domestic chickens, Gallus domesticus, for comparison. All individuals were sexed successfully with high reproducibility. We conclude that this PCR-based test with feathers as the DNA source is a reliable sexing method for H. leucogaster. This sexing technique is objective and non-invasive and could be used to test sex ratio theories, as well as to help improve conservation and management actions for captive breeding program of this species in Malaysia.
The influence of physical and chemical parameters on the abundance and diversity of chironomids was studied in six rivers with moderate to highly polluted water in the Juru River Basin. The rivers: Ceruk Tok Kun (CTKR) as reference site, and polluted rivers of Pasir (PR), Juru (JR), Permatang Rawa (PRR), Ara (AR) and Kilang Ubi (KUR) were sampled over a period of five months (November 2007-March 2008). Nine chirnomid species: Chironimus kiiensis, C. javanus, Polypedilum trigonus, Microchironomus sp., Dicrotendipes sp., Tanytarsus formosanus, Clinotanypus sp., Tanypus punctipennis and Fittkauimyia sp. were identified. Assessment of their relationships with several environmental parameters was performed using the canonical correspondence analysis (CCA). Tanytarsus formosanus was the most dominant in the relatively clean CTKR and moderately polluted JR with mean densities of 19.66 and 25.32 m(-2), respectively while C. kiiensis was abundant in more polluted rivers. Tanytarsus formosanus, Dicrotendipes sp. and Microchironomus sp. were grouped under moderate to high water temperature, total organic matter (TOM), total suspended solids (TSS), velocity, pH, phosphates and sulphates. However, Tanypus punctipennis, Fittkauimyia sp., and Clinotanypus sp. were associated with high contents of river sediment such as TOM, Zn and Mn and water ammonium-N and nitrate-N and they were associated with higher dissolved oxygen (DO) content in the water. Chironomus kiiensis, C. javanus and P. trigonus showed positive relationships with TOM, ammonium-N and nitrate-N as well as trace metals of Zn, Cu and Mn. These three species could be considered as tolerant species since they have the ability to survive in extreme environmental conditions with low DO and high concentrations of pollutants. Based on the water parameter scores in all rivers, the highest diversity of chironomid larvae was reported in CTKR. With higher concentrations of organic and/or inorganic pollutants as reported in PPR, KUR and AR, the chironomid larval diversity decreased, and the abundance of tolerant species, mainly Chironomus spp., increased.
This study used a two-phase feeding trial to determine the influence of selected dietary prebiotics and probiotics on growth performance, feed utilisation, and morphological changes in snakehead (Channa striata) fingerlings as well as the duration of these effects over a post-experimental period without supplementation. Triplicate groups of fish (22.46 ±0.17 g) were raised on six different treatment diets: three prebiotics (0.2% β-glucan, 1% galacto-oligosaccharides [GOS], 0.5% mannan-oligosaccharides [MOS]), two probiotics (1% live yeast [Saccharomyces cerevisiae] and 0.01% Lactobacillus acidophilus [LBA] powder) and a control (unsupplemented) diet; there were three replicates for each treatment. All diets contained 40% crude protein and 12% crude lipid. Fish were fed to satiation three times daily. No mortalities were recorded during Phase 1; however, 14% mortality was documented in the control and prebiotic-amended fish during Phase 2. At the end of Phase 1, growth performance and feed utilisation were significantly higher (p<0.05) in the LBA-treated fish, followed by live yeast treatment, compared with all other diets tested. The performance of fish on the three prebiotic diets were not significantly different from one another but was significantly higher than the control diet. During Phase 2 (the post-feeding phase), fish growth continued until the 6th week for the probiotic-based diets but levelled off after four weeks for the fish fed the prebiotic diets. The feed conversion ratio (FCR) was higher in all treatments during the post-feeding period. The hepatosomatic index (HSI) did not differ significantly among the tested diets. The visceral somatic index (VSI) and intraperitoneal fat (IPF) were highest in the LBA-based diet and the control diet, respectively. The body indices were significantly different (p<0.05) between Phases 1 and 2. This study demonstrates that probiotic-based diets have a more positive influence on the growth, feed utilisation, and survival of C. striata fingerlings compared with supplementation with prebiotics.
A newly discovered, diminutive, cave-dwelling, lowland species of the colubrid snake genus Lycodon Boie is described from a limestone cave along the Thai-Malaysian border in the state of Perlis, northwestern Peninsular Malaysia. Lycodon cavernicolus sp. nov. is most closely related to L. butleri Boulenger, an endemic, upland, forest-dwelling species from Peninsular Malaysia of the fasciatus group but is separated from L. butleri and all other species of the L. fasciatus group and the closely related L. ruhstrati group by having the combination of 245 (male) and 232 (female) ventral scales; 113 (male) and 92 (female) paired, subcaudal scales; a single precloacal plate; nine or 10 supralabials; 10 or 11 infralabials; a maximum total length of 508 mm (female); a relative tail length of 0.25-0.27; an immaculate venter in juveniles and dark-brown, posterior, ventral scale margins in adults; and dorsal and caudal bands in juveniles white. The discovery of L. cavernicolus sp. nov. adds to a rapidly growing list of newly discovered reptiles from karst regions and limestone forests of Peninsular Malaysia, underscoring the fact that these areas should be studied before they are quarried as they harbor a significant portion of the Peninsular Malaysia's herpetological diversity.
Mitochondrial cytochrome oxidase subunit I (COI) gene was utilized to assess the population genetics of the commercially important black scar oyster, Crassostrea iredalei among 11 populations throughout the west and east coasts Peninsular Malaysia and Sabah (Malaysian Borneo). Overall, populations of C. iredalei demonstrated low nucleotide diversity π (0.000-0.004) and low-to-high haplotype diversity h (0.000-0.795) levels. Genetic structuring was detected between the Peninsular Malaysia and Sabah populations as revealed by the FST analysis. However, the COI gene analyses showed minimal and non-significant (p > 0.05) population differentiation within the east and west coasts Peninsular Malaysia and Sabah regions. This was attributed to both high larval dispersal along the east and west coasts and human-driven spat translocation between the two coastlines due to C. iredalei cultivation practices. Phylogeographic relationships inferences were also conducted to further support these hypotheses. The neutrality and mismatch distribution analyses suggested that C. iredalei had experienced a/several bottleneck event(s), followed by population expansion. The molecular information obtained from this study could be incorporated in a pragmatic aquaculture management strategy of wild broodstock and the hatchery lines of C. iredalei in Malaysia.
A total of 30 specimens belonging to five species, namely; Cryptozona siamensis, Sarika resplendens and Sarika sp. from the family Ariophantidae as well as Quantula striata and Quantula sp. from the family Dyakiidae were collected from the Langkawi Island in Northern Peninsular Malaysia. All specimens were identified through comparisons of shell morphology and amplification of a 500 bp segment of the 16S rRNA mtDNA gene. To assess phylogenetic insights, the sequences were aligned using ClustalW and phylogenetic trees were constructed. The analyses showed two major lineages in both Maximum Parsimony and Neighbour Joining phylogenetic trees. Each putative taxonomic group formed a monophyletic cluster. Our study revealed low species and intraspecies genetic diversities based on the 16S rRNA gene sequences. Thus, this study has provided an insight of land snail diversity in populations of an island highly influenced by anthropogenic activities through complementary use of shell morphological and molecular data.
Abundance and diversity of benthic macroinvertebrates as well as physico-chemical parameters were investigated in five rivers of the Juru River Basin in northern Peninsula Malaysia: Ceruk Tok Kun River (CTKR), Pasir River (PR), Permatang Rawa River (PRR), Kilang Ubi River (KUR), and Juru River (JR). The physico-chemical parameters and calculated water quality index (WQI) were significantly different among the investigated rivers (ANOVA, P<0.05). The WQI classified CTKR, PR, and JR into class III (slightly polluted). However, PRR and KUR fell into class IV (polluted). High diversity and abundance of macroinvertebrates, especially the intolerant taxa, Ephemeroptera, Plecoptera, and Trichoptera, were observed in the least polluted river, CTKR. Decreasing abundance of macroinvertebrates followed the deterioration of river water quality with the least number of the most tolerant taxa collected from PR. On the basis of composition and sensitivity of macroinvertebrates to pollutants in each river, the highest Biological Monitoring Working Party (BMWP) index score of 93 was reported in CTKR (good water quality). BMWP scores in PRR and JR were 38.7 and 20.1, respectively, classifying both of them into "moderate water quality" category. Poor water quality was reported in PR and KUR. The outcome of the multivariate analysis (CCA) was highly satisfactory, explaining 43.32% of the variance for the assemblages of macroinvertebrates as influenced by 19 physical and chemical variables. According to the CCA model, we assert that there were three levels of stresses on macroinvertebrate communities in the investigated rivers: Level 1, characterized of undisturbed or slightly polluted as in the case of CTKR; Level 2, characterized by a lower habitat quality (the JR) compared to the CTKR; and Level 3 showed severe environmental stresses (PRR, PR, and KUR) primarily contributed by agricultural, industrial, and municipal discharges.
The genotoxic effects of increasing concentrations (below lethal concentration [LC₅₀]) of cadmium ([Cd] 0.1, 1 and 10 mg/L), copper ([Cu] 0.2, 2 and 20 mg/L) and zinc ([Zn] 0.5, 5 and 50 mg/L) on Chironomus kiiensis were evaluated using alkaline comet assay after exposure for 24 h. Both the tail moment and the olive tail moment showed significant differences between the control and different concentrations of Cd, Cu and Zn (Kruskal-Wallis, p < 0.05). The highest concentration of Cd was associated with higher DNA damage to C. kiiensis larvae compared with Cu and Zn. The potential genotoxicity of these metals to C. kiiensis was Cd > Cu > Zn.
Morphological deformities in parts of the head capsule of Chironomus spp. larvae inhabiting three polluted rivers (Permatang Rawa [PRR], Pasir [PR], and Kilang Ubi [KUR]) in the Juru River Basin, northeastern peninsular Malaysia, were studied. Samples of the fourth-instar larvae at one location in each river were collected monthly from November 2007 to March 2008 and examined for deformities of the mentum, antenna, mandible, and epipharyngis. At each sample location, in situ measurements of water depth, river width, water pH, dissolved oxygen, and water temperature were made. Samples of river water and benthic sediments were also collected monthly from each larval sample location in each river and taken to the laboratory for appropriate analysis. Total suspended solids (TSSs), ammonium-N, nitrate-N, phosphate-P, chloride, sulfate, and aluminum content in water were analyzed. Total organic matter and nonresidual metals in the sediment samples were also analyzed. Among the three rivers, the highest mean deformity (47.17%) was recorded in larvae collected from KUR that received industrial discharges from surrounding garment and rubber factories, followed by PRR (33.71%) receiving primarily residues of fertilizers and pesticides from adjacent rice fields, and PR (30.34%) contaminated primarily by anthropogenic wastes from the surrounding residential areas. Among the various head capsule structures, deformity of the mentum was strongly reflective of environmental stress and amounted to 27.9, 20.87, and 30.19% in the PRR, PR, and KUR, respectively. Calculated Lenat's toxic score index satisfactorily explained the influence of prevailing environmental variables on the severity of mentum deformities. Redundancy analysis and forward selection selected TSSs, sediment Zn, Mn, Cu, and Ni, and water pH, dissolved oxygen, water temperature, total organic matter, nitrate-N, chloride, phosphate-P, ammonium-N, sulfate, and aluminum as parameters that significantly affected some proportion of deformities. The total deformities correlated closely with deformities of mentum but only weakly with deformities in other parts of head. The total deformity incidence was strongly correlated with high contents of sediment Mn and Ni. The mentum and epipharyngis deformities incidence was highly correlated with an increase of TSSs, total aluminum, and ammonium-N and a decrease in pH and dissolved oxygen.
DNA barcodes, typically focusing on the cytochrome oxidase I gene (COI) in many animals, have been used widely as a species-identification tool. The ability of DNA barcoding to distinguish species from a range of taxa and to reveal cryptic species has been well documented. Despite the wealth of DNA barcode data for fish from many temperate regions, there are relatively few available from the Southeast Asian region. Here, we target the marine fish Family Carangidae, one of the most commercially-important families from the Indo-Malay Archipelago (IMA), to produce an initial reference DNA barcode library.
Phylogeographic patterns and population structure of the pelagic Indian mackerel, Rastrelliger kanagurta were examined in 23 populations collected from the Indonesian-Malaysian Archipelago (IMA) and the West Indian Ocean (WIO). Despite the vast expanse of the IMA and neighbouring seas, no evidence for geographical structure was evident. An indication that R. kanagurta populations across this region are essentially panmictic. This study also revealed that historical isolation was insufficient for R. kanagurta to attain migration drift equilibrium. Two distinct subpopulations were detected between the WIO and the IMA (and adjacent populations); interpopulation genetic variation was high. A plausible explanation for the genetic differentiation observed between the IMA and WIO regions suggest historical isolation as a result of fluctuations in sea levels during the late Pleistocene. This occurrence resulted in the evolution of a phylogeographic break for this species to the north of the Andaman Sea.
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.
Chironomus javanus (Kieffer) and Chironomus kiiensis Tokunaga were redescribed from materials collected from a rice field in Pulau Pinang, Malaysia. The larvae can only be distinguished after careful preparation and examination using a compound microscope, but the pupae were not useful to differentiate C. javanus from C. kiiensis. The adult specimens showed clear body and wing characteristics for rapid and accurate identification.
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.