Since 2010, several duck Tembusu viruses (DTMUVs) have been isolated from infected ducks in China, and these virus strains have undergone extensive variation over the years. Although the infection rate is high, the mortality rate is usually relatively low-~5%-30%; however, since fall 2019, an infectious disease similar to DTMUV infection but with a high mortality rate of ~50% in goslings has been prevalent in Anhui Province, China. The present study identified a new Tembusu virus, designated DTMUV/Goose/China/2019/AQ-19 (AQ-19), that is believed to be responsible for the noticeably high mortality in goslings. To investigate the genetic variation of this strain, its entire genome was sequenced and analysed for specific variations, and goslings and mice were challenged with the isolated virus to investigate its pathogenicity. The AQ-19 genome shared only 94.3%-96.9% and 90.9% nucleotide identity with other Chinese and Malaysian DTMUVs, respectively; however, AQ-19 has high homology with Thailand DTMUVs (97.2%-98.1% nucleotide identity). Phylogenetic analysis of the E gene revealed that AQ-19 and most of Thailand DTMUVs form a branch separate from any of the previously reported DTMUV strains in China. After the challenge, some goslings and mice showed typical clinical signs of DTMUV, particularly severe neurological dysfunction. AQ-19 has high virulence in goslings and mice, resulting in 60% and 70% mortality through intramuscular and intracerebral routes, respectively. Pathological examination revealed severe histological lesions in the brain and liver of the infected goslings and mice. Taken together, these results demonstrated the emergence of a novel Tembusu virus with high virulence circulating in goslings in China for the first time, and our findings highlight the high genetic diversity of DTMUVs in China. Further study of the pathogenicity and host range of this novel Tembusu virus is particularly important.
Papaya ringspot virus (PRSV) is a plant virus transmitted by aphids that has spread throughout many countries, including Malaysia, causing yield losses and economic impacts to the papaya industry worldwide. PRSV infection in papaya-distinctive ring-shaped patterns on papaya leaves resulted in stunted growth and reduced fruit quality. Management strategies such as the use of resistant varieties, cultural practices, and vector control are employed to mitigate the spread of PRSV. However, the evolution of new virus strains and the uncertainties posed by climate change pose ongoing challenges for the management of PRSV worldwide. Therefore, in this present study, we aim to confirm the presence of PRSV in symptomatic papaya leaves, to depict the current status of PRSV in Malaysia. Using reverse-transcription PCR (RT-PCR) targeting PRSV partial nuclear inclusion b protein (NIb) and coat protein (CP), 13 out of 40 papaya leaves collected were found positive for the PRSV strain-P (PRSV-P). Nucleotide analysis revealed a high similarity with strains from Taiwan and India, showing 96.83%, 97.03%, and 97.03% identity with the Taiwan strains (DQ340771, AY027810) and the India strain (KJ755852), respectively. Compared to the CP gene of Malaysian isolates reported in 2016 (EU082207), several nonsynonymous mutations have been discovered suggesting genetic diversity within the PRSV population in Malaysia. Overall, this study confirms the current circulation of PRSV infection in Malaysia since it was first identified in Johore in 1991. The re-occurrence of PRSV-P in this study highlights the need for continuous monitoring and targeted management strategies to prevent the further spread of PRSV-P in Malaysia.
The following nine elongate unpatterned muraenid species of the subfamily Muraeninae, including one new species, are recognized from Taiwan and adjacent waters: Gymnothorax albimarginatus (Temminck & Schlegel), G. dorsalis Seale, G. melanosomatus Loh, Shao & Chen, G. phasmatodes (Smith), G. prolatus Sasaki & Amaoka, G. sagmacephalus Böhlke, Pseudechidna brummeri (Bleeker), Strophidon sathete (Hamilton) and G. pseudomelanosomatus new species, described from two specimens. This new moray eel is distinguished from its similar species, G. melanosomatus, by the following features: grey brown body (vs. black), snout length 20.5% (vs. 17.8%) of head length, smaller eye diameter 8.2% (vs. 10.0%) of head length; preanal length 49.5% (vs. 58.5%) total length, and preanal vertebrae 89-89 (vs. 105-109). Phylogenetic relationships of the nine species were examined using nucleotide sequence data from partial sequences of mitochondrial ND5 gene (600 bp), and seven species form COI (600 bp). The genetic analyses suggest that G. pseudomelanosomatus is distinct from G. melanosomatus and the other six species of Gymnothorax. Morphological features and mitogenetic affinities strongly suggest that "G." dorsalis should be placed in Strophidon rather than in Gymnothorax. The results also suggest that employment of ND5 and COI gene sequences are rather useful for identification of species and for obtaining reasonable insights into the phylogeny of the muraenid species.
Schistosomiasis is a neglected tropical disease that affects more than 200 million people worldwide. The main disease-causing agents, Schistosoma japonicum, S. mansoni and S. haematobium, are blood flukes that have complex life cycles involving a snail intermediate host. In Asia, S. japonicum causes hepatointestinal disease (schistosomiasis japonica) and is challenging to control due to a broad distribution of its snail hosts and range of animal reservoir hosts. In China, extensive efforts have been underway to control this parasite, but genetic variability in S. japonicum populations could represent an obstacle to eliminating schistosomiasis japonica. Although a draft genome sequence is available for S. japonicum, there has been no previous study of molecular variation in this parasite on a genome-wide scale. In this study, we conducted the first deep genomic exploration of seven S. japonicum populations from mainland China, constructed phylogenies using mitochondrial and nuclear genomic data sets, and established considerable variation between some of the populations in genes inferred to be linked to key cellular processes and/or pathogen-host interactions. Based on the findings from this study, we propose that verifying intraspecific conservation in vaccine or drug target candidates is an important first step toward developing effective vaccines and chemotherapies against schistosomiasis.
Taxonomic status of fanged frogs from the Peninsular Malaysia, previously assigned to Limnonectes kuhlii, is assessed using genetic and morphological approaches. Phylogenetic relationships inferred from sequences of the mitochondrial and nuclear genes revealed that the fanged frogs from the Peninsula form a monophyletic group and are clearly divergent from other species previously, or even now, assigned to L. kuhlii from Mainland Southeast Asia. In both mtDNA and nuDNA phylogeny, the Malay Peninsula clade diverges into two lineages, one from north (Larut Hill, Perak, and Hulu Terengganu, Terengganu) and another from south (Genting Highlands, Pahang, and Gombak, Selangor). These lineages are separated by large genetic distances, comparable with those observed between some other species of L. kuhlii-like frogs. Although the two lineages are very similar morphologically, they are distinguishable in several morphological traits and are considered heterospecific. We therefore describe them as L. utara sp. nov. and L. selatan sp. nov. These new species differ from all other species of kuhlii-like frogs from Mainland Southeast Asia by the surface of tibia, which is densely covered by large warts.
Freshwater eels have fascinated biologists for centuries due to the spectacular long-distance migrations between the eels' freshwater habitats and their spawning areas far out in the ocean and the mysteries of their ecology. The spawning areas of Atlantic eels and Japanese eel were located far offshore in the Atlantic Ocean and the Pacific Ocean, respectively, and their reproduction took place thousands of kilometers away from their growth habitats. Phylogenetic studies have revealed that freshwater eels originated in the Indonesian region. However, remarkably little is known about the life histories of tropical freshwater eels despite the fact that tropical eels are key to understanding the nature of primitive forms of catadromous migration. This study found spawning-condition tropical freshwater eels in Lake Poso, central Sulawesi, Indonesia, with considerably high gonadosomatic index values and with histologically fully developed gonads. This study provides the first evidence that under certain conditions, freshwater eels have conditions that are immediately able to spawn even in river downstream. The results suggest that, in contrast to the migrations made by the Atlantic and Japanese eels, freshwater eels originally migrated only short distances of <100 kilometers to local spawning areas adjacent to their freshwater growth habitats. Ancestral eels most likely underwent a catadromous migration from local short-distance movements in tropical coastal waters to the long-distance migrations characteristic of present-day temperate eels, which has been well established as occurring in subtropical gyres in both hemispheres.
A well-supported and well-resolved phylogeny based on a concatenated data set from one mitochondrial and two nuclear genes, six morphological characters, and nine color pattern characters for 44 of the 50 species of the Southeast Asian Rock Geckos (genus Cnemaspis Strauch, 1887) is consistent with the previous taxonomy of Cnemaspis based solely on morphology and color pattern. Cnemaspis is partitioned into four major clades that collectively contain six species groups. The monophyly of all clades and species groups is strongly supported and they are parapatrically distributed across well-established, biogeographical regions ranging from southern Vietnam westward through southern Indochina, southward through the Thai-Malay Peninsula, then eastward to Borneo. Eight new species (Cnemaspis omari sp. nov. from the Thai-Malaysian border; C. temiah sp. nov. from Cameron Highlands, Pahang, Malaysia; C. stongensis sp. nov. from Gunung Stong, Kelantan, Malaysia; C. hangus sp. nov. from Bukit Hangus, Pahang, Malaysia; C. sundagekko sp. nov. from Pulau Siantan, Indonesia; C. peninsularis sp. nov. from southern Peninsular Malaysia and Singapore, and C. mumpuniae sp. nov. and C. sundainsula sp. nov. from Pulau Natuna Besar, Indonesia) are described based on morphology and color pattern and all but C. sundagekko sp. nov. are included in the phylogenetic analyses. Cnemaspis kendallii is polyphyletic and a composite of six species. An updated taxonomy consistent with the phylogeny is proposed for all 50 species and is based on 25 morphological and 53 color pattern characters scored across 594 specimens. Cladogenetic events and biogeographical relationships within Cnemaspis were likely influenced by this group's low vagility and the cyclical patterns of geographical and environmental changes in Sundaland over the last 25 million years and especially within the last 2.5 million years. The phylogeny indicates that nocturnality, diurnality, substrate preferences, and the presence of ocelli in the shoulder regions have evolved independently multiple times.
Alexandrium ostenfeldii (Paulsen) Balech and Tangen and A. peruvianum (Balech and B.R. Mendiola) Balech and Tangen are morphologically closely related dinoflagellates known to produce potent neurotoxins. Together with Gonyaulax dimorpha Biecheler, they constitute the A. ostenfeldii species complex. Due to the subtle differences in the morphological characters used to differentiate these species, unambiguous species identification has proven problematic. To better understand the species boundaries within the A. ostenfeldii complex we compared rDNA data, morphometric characters and toxin profiles of multiple cultured isolates from different geographic regions. Phylogenetic analysis of rDNA sequences from cultures characterized as A. ostenfeldii or A. peruvianum formed a monophyletic clade consisting of six distinct groups. Each group examined contained strains morphologically identified as either A. ostenfeldii or A. peruvianum. Though key morphological characters were generally found to be highly variable and not consistently distributed, selected plate features and toxin profiles differed significantly among phylogenetic clusters. Additional sequence analyses revealed a lack of compensatory base changes in ITS2 rRNA structure, low to intermediate ITS/5.8S uncorrected genetic distances, and evidence of reticulation. Together these data (criteria currently used for species delineation in dinoflagellates) imply that the A. ostenfeldii complex should be regarded a single genetically structured species until more material and alternative criteria for species delimitation are available. Consequently, we propose that A. peruvianum is a heterotypic synonym of A. ostenfeldii and this taxon name should be discontinued.
Although recent molecular studies have indicated the presence of a number of distinct species within the Caulerpa racemosa-peltata complex, due to the difficulties presented by high levels of phenotypic plasticity and the large number of synonyms, infra-specific taxa, and names of uncertain affinity, taxonomic proposals are yet to be made. In this study, we aimed to resolve the taxonomy of the complex and provide an example of how historical nomenclature can best be integrated into molecular based taxonomies. We accomplished this by first determining the number of genetic species within our globally sampled data set through a combination of phylogenetic and species-delimitation approaches of partial elongation factor TU and RUBISCO large subunit gene sequences. Guided by these results, comparative morphological examinations were then undertaken to gauge the extent of phenotypic plasticity within each species, as well as any morphological overlap between them. Our results revealed the presence of 11 distinct species within the complex, five of which showed high levels of phenotypic plasticity and partial overlap with other species. On the basis of observations of a large number of specimens, including type specimens/descriptions, and geographic inferences, we were able to confidently designate names for the lineages. Caulerpa peltata, C. imbricata and C. racemosa vars. laetevirens, occidentalis and turbinata were found to represent environmentally induced forms of a single species, for which the earlier-described C. chemnitzia, previously regarded as a synonym of C. racemosa var. turbinata, is reinstated. C. cylindracea, C. lamourouxii, C. macrodisca, C. nummularia and C. oligophylla are also reinstated and two new species, C. macra stat. nov. and C. megadisca sp. nov., are proposed.
We record a tree frog of the genus Chiromantis for the first time from outside the Southeast Asian continent and describe it as a new species, Chiromantis inexpectatus. The new species from the Malaysian state of Sabah, Borneo, is a small-sized Chiromantis (male snout-vent length ca. 22 mm), and is distinguished from all other members of the genus by the combination of the following morphological characteristics: dark stripes absent, but dark spots present on dorsum; a dark-brown lateral band present from snout tip to half of body, bordered ventrally by white stripe; third and fourth fingers less than half webbed; third finger disk wider than tympanum diameter; and inner metatarsal tubercle present. Significance of findings of this species from Borneo Island, as well as phylogeny and breeding habit of the genus Chiromantis, are briefly discussed.
Coolia is a widespread and ecologically important genus of benthic marine dinoflagellates found in tropical regions. Historically, there has been taxonomic confusion about the taxonomy and toxicity of this group. The goal of this study was to resolve morphological questions concerning Coolia tropicalis and determine the taxonomic identity of the Australian Coolia isolate which has been reported to produce cooliatoxins. To accomplish this, the morphology of tropical strains from Belize (the type locality of C. tropicalis), Malaysia, Indonesia, and Australia were examined and compared to published reports. The morphological analysis showed that C. tropicalis differs from the original description in that it has a slightly larger size (35-47 μm long by 30-45 μm wide versus 23-40 μm long by 25-39 μm wide), and the shape of fourth apical plate, and the length of Po plate (7.4-12 μm versus 7 μm). Based on both morphology and phylogenetic analysis using LSU D1- D3 rDNA sequences, the clones of C. tropicalis from Malaysia, Indonesia, and Belize were found to form a monophyletic clade within the genus. The strain producing cooliatoxin was found to be C. tropicalis, not Coolia monotis as originally assumed. To explore the factors influencing the growth of Coolia species, the growth rates of C. tropicalis and Coolia malayensis were determined at different temperatures and salinities. Both species tolerated a wide range of temperatures, but cannot survive at temperatures <20°C or >35°C. C. monotis, the dominant species reported in the literature, probably does not produce toxins.
Genetic diversity is prerequisite for any crop improvement program as it helps in the development of superior recombinants. Fifty Malaysian upland rice accessions were evaluated for 12 growth traits, yield and yield components. All of the traits were significant and highly significant among the accessions. The higher magnitudes of genotypic and phenotypic coefficients of variation were recorded for flag leaf length-to-width ratio, spikelet fertility, and days to flowering. High heritability along with high genetic advance was registered for yield of plant, days to flowering, and flag leaf length-to-width ratio suggesting preponderance of additive gene action in the gene expression of these characters. Plant height showed highly significant positive correlation with most of the traits. According to UPGMA cluster analysis all accessions were clustered into six groups. Twelve morphological traits provided around 77% of total variation among the accessions.
A taxonomic study of the genus Padina from Japan, Southeast Asia, and Hawaii based on morphology and gene sequence data (rbcL and cox3) resulted in the recognition of four new species, that is, Padina macrophylla and Padina ishigakiensis from Ryukyu Islands, Japan; Padina maroensis from Hawaii; and Padina usoehtunii from Myanmar and Thailand. All species are bistratose and morphologically different from one another as well as from any known taxa by a combination of characters relating to degree of calcification; the structure, position, and arrangement of hairlines (HLs) and reproductive sori; and the presence or absence of rhizoid-like groups of hairs and an indusium. Molecular phylogenetic analyses demonstrated a close relationship between P. ishigakiensis, P. macrophylla, P. maroensis, and Padina australis Hauck. The position of P. usoehtunii, however, was not fully resolved, being either sister to a clade comprising the other three new species and P. australis in the rbcL tree or more closely related to a clade comprising several other recently described species in the cox3 tree. The finding of the four new species demonstrates high species diversity particularly in southern Japan. The following characters were first recognized here to be useful for species delimitation: the presence or absence of small rhizoid-like groups of hairs on the thallus surface, structure and arrangement of HLs on both surfaces either alternate or irregular, and arrangement of the alternating HLs between both surfaces in equal or unequal distance. The evolutionary trajectory of these and six other morphological characters used in species delineation was traced on the phylogenetic tree.
This note redescribes M. borneensis and describes M. traubi n. sp. based on the known specimens from the two sampling localities (holotype from Mount Murud and further recorded from Mount Kinabalu by Traub). The clearly allied, but clearly distinct species for the two localities, were recorded.
Anew hydrophilid genus Chimaerocyon gen. nov. containing two species, C. shimadai sp. nov. (Malaysia: Pahang) and C. sumatranus sp. nov. (Indonesia: Sumatra), is described. Specimens of C. shimadai were collected from brood cells in anest of Pheidole singaporensis Özdikmen, 2010. The biology of C. sumatranus remains unknown. A molecular phylogeny based on four genes (cox1, cox2, 18S and 28S) supports the placement of the genus as deeply nested within the Cercyon-group of the tribe Megasternini. This position is supported by the subdistal position of the median spur in the hind wing (unique to Megasternini) and the presence of sucking disc on male maxilla (unique for Megastemini+Sphaeridiini). The remaining external morphology differs substantially from other representatives of Megasternini. The hypothesis that the aberrant morphology of Chimaerocyon gen. nov. is a consequence of myrmecophily is discussed.
PHA synthase (PhaC) is the key enzyme in the production of biodegradable plastics known as polyhydroxyalkanoate (PHA). Nevertheless, most of these enzymes are isolated from cultivable bacteria using traditional isolation method. Most of the microorganisms found in nature could not be successfully cultivated due to the lack of knowledge on their growth conditions. In this study, a culture-independent approach was applied. The presence of phaC genes in limestone soil was screened using primers targeting the class I and II PHA synthases. Based on the partial gene sequences, a total of 19 gene clusters have been identified and 7 clones were selected for full length amplification through genome walking. The complete phaC gene sequence of one of the clones (SC8) was obtained and it revealed 81% nucleotide identity to the PHA synthase gene of Chromobacterium violaceum ATCC 12472. This gene obtained from uncultured bacterium was successfully cloned and expressed in a Cupriavidus necator PHB(-)4 PHA-negative mutant resulting in the accumulation of significant amount of PHA. The PHA synthase activity of this transformant was 64 ± 12 U/g proteins. This paper presents a pioneering study on the discovery of phaC in a limestone area using metagenomic approach. Through this study, a new functional phaC was discovered from uncultured bacterium. Phylogenetic classification for all the phaCs isolated from this study has revealed that limestone hill harbors a great diversity of PhaCs with activities that have not yet been investigated.
Prophages of Helicobacter pylori, a bacterium known to co-evolve in the stomach of its human host, were recently identified. However, their role in the diversity of H. pylori strains is unknown. We demonstrate here and for the first time that the diversity of the prophage genes offers the ability to distinguish between European populations, and that H. pylori prophages and their host bacteria share a complex evolutionary history. By comparing the phylogenetic trees of two prophage genes (integrase and holin) and the multilocus sequence typing (MLST)-based data obtained for seven housekeeping genes, we observed that the majority of the strains belong to the same phylogeographic group in both trees. Furthermore, we found that the Bayesian analysis of the population structure of the prophage genes identified two H. pylori European populations, hpNEurope and hpSWEurope, while the MLST sequences identified one European population, hpEurope. The population structure analysis of H. pylori prophages was even more discriminative than the traditional MLST-based method for the European population. Prophages are new players to be considered not only to show the diversity of H. pylori strains but also to more sharply define human populations.
In this study, we sequenced the genome of Pandoraea pnomenusa RB38 using Pacific Biosciences RSII (PacBio) Single Molecule Real Time (SMRT) sequencing technology. A pair of cognate luxI/R homologs was identified where the luxI homolog, ppnI, was found adjacent to a luxR homolog, ppnR1. An additional orphan luxR homolog, ppnR2, was also discovered. Multiple sequence alignment and phylogenetic analysis revealed that ppnI is an N-acyl homoserine lactone (AHL) synthase gene that is distinct from those of the nearest phylogenetic neighbor viz. Burkholderia spp. High resolution tandem mass spectrometry (LC-MS/MS) analysis showed that Escherichia coli BL21 harboring ppnI produced a similar AHL profile (N-octanoylhomoserine lactone, C8-HSL) as P. pnomenusa RB38, the wild-type donor strain, confirming that PpnI directed the synthesis of AHL in P. pnomenusa RB38. To our knowledge, this is the first documentation of the luxI/R homologs of the genus Pandoraea.
Wolbachia is an alpha-proteobacterial symbiont widely distributed in arthropods. Since the identification of Wolbachia in certain animal-parasitic nematodes (the Onchocercidae or filariae), the relationship between arthropod and nematode Wolbachia has attracted great interest. The obligate symbiosis in filariae, which renders infected species susceptible to antibiotic chemotherapy, was held to be distinct from the Wolbachia-arthropod relationship, typified by reproductive parasitism. While co-evolutionary signatures in Wolbachia-arthropod symbioses are generally weak, reflecting horizontal transmission events, strict co-evolution between filariae and Wolbachia has been reported previously. However, the absence of close outgroups for phylogenetic studies prevented the determination of which host group originally acquired Wolbachia. Here, we present the largest co-phylogenetic analysis of Wolbachia in filariae performed to date including: (i) a screening and an updated phylogeny of Wolbachia; (ii) a co-phylogenetic analysis; and (iii) a hypothesis on the acquisition of Wolbachia infection. First, our results show a general overestimation of Wolbachia occurrence and support the hypothesis of an ancestral absence of infection in the nematode phylum. The accuracy of supergroup J is also underlined. Second, although a global pattern of coevolution remains, the signal is derived predominantly from filarial clades associated with Wolbachia in supergroups C and J. In other filarial clades, harbouring Wolbachia supergroups D and F, horizontal acquisitions and secondary losses are common. Finally, our results suggest that supergroup C is the basal Wolbachia clade within the Ecdysozoa. This hypothesis on the origin of Wolbachia would change drastically our understanding of Wolbachia evolution.
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.