Southeast Asian Species Research Articles

Pleistocene events and present environmental factors have shaped the phylogeography of the intertidal limpetCellana toreuma(Reeve, 1855) (Gastropoda: Nacellidae) in Southeast Asia and China

High levels of biodiversity and phylogeographic structure in marine species in Southeast Asia are strongly linked to Quaternary sea-level fluctuations and complex oceanographic conditions. Cellana toreuma is a common limpet on intertidal rocky shores and is widely distributed in the Western Pacific. Analyses of partial mitochondrial COI gene sequences from Southeast Asia, combined with previously published sequences from East Asia and Indonesia, revealed the existence of five well-supported clades with high genetic divergences (between 1.4 and 7.6%), namely the East Asia clade, the eastern Southeast Asia clade, the western Southeast Asia clade, the Pelabuan Ratu (Java) clade and the Ogasawara clade. The geographical distribution of the five clades is likely related to the history of glaciations and rapid postglacial population expansions. Analyses of pairwise ΦST and hierarchical analysis of molecular variance shows significant population structure among collections in East and Southeast Asia. These results suggest that historical events have had strong effects on the phylogeographic structure of C. toreuma. In addition, present environmental factors, such as unsuitable habitats and ocean currents, have also affected the genetic footprints of past environments.

Polyphyly of Asian Tree Toads, Genus Pedostibes Günther, 1876 (Anura: Bufonidae), and the Description of a New Genus from Southeast Asia.

The Asian Tree Toad genus Pedostibes, as currently understood, exhibits a conspicuously disjunct distribution, posing several immediate questions relating to the biogeography and taxonomy of this poorly known group. The type species, P. tuberculosus and P. kempi, are known only from India, whereas P. hosii, P. rugosus, and P. everetti are restricted to Southeast Asia. Several studies have shown that these allopatric groups are polyphyletic, with the Indian Pedostibes embedded within a primarily South Asian clade of toads, containing the genera Adenomus, Xanthophryne, and Duttaphrynus. Southeast Asian Pedostibes on the other hand, are nested within a Southeast Asian clade, which is the sister lineage to the Southeast Asian river toad genus Phrynoidis. We demonstrate that Indian and Southeast Asian Pedostibes are not only allopatric and polyphyletic, but also exhibit significant differences in morphology and reproductive mode, indicating that the Southeast Asian species’ are not congeneric with the true Pedostibes of India. As a taxonomic solution, we describe a new genus, Rentapia gen. nov. to accommodate the Southeast Asian species.

Description of two final stadium Onychogomphus larvae from Thailand(Odonata: Gomphidae).

The final stadium larvae of Onychogomphus castor Lieftinck and O. duaricus Fraser are described and illustrated for the first time based on reared specimens from Thailand. The taxonomic characteristics of the larvae of the genus Onychogomphus are discussed and summarized. The larva of O. castor differs from other Southeast Asian species in having distinct mid-dorsal spines on S2-9, divergent wing pads reaching S5, and lateral spines on S6-9. The larva of O. duaricus has a weakly swollen third antennal segment, with short blunt mid-dorsal spines on S2-9, divergent wing sheaths reaching the middle of S4, and lateral spines present on S7-9.

The Australasian frog family Ceratobatrachidae in China, Myanmar and Thailand: discovery of a new Himalayan forest frog clade.

In an effort to study the systematic affinities and specieslevel phylogenetic relationships of the enigmatic anurans variably assigned to the genera Ingerana or Limnonectes (family Dicroglossidae), we collected new molecular sequence data for five species including four Himalayan taxa, Limnonectes xizangensis, Lim. medogensis, Lim. alpina, [corrected] Ingerana borealis and one southeast Asian species, I. tasanae, and analyzed these together with data from previous studies involving other ostensibly related taxa. Our surprising results demonstrate unequivocally that Lim. xizangensis, Lim. medogensis and Lim. alpina [corrected] form a strongly supported clade, the sister-group of the family Australasian forest frog family Ceratobatrachidae. This discovery requires an expansion of the definition of Ceratobatrachidae and represents the first record of this family in China. These three species are distinguished from the species of Ingerana and Limnonectes by the: (1) absence of interdigital webbing of the foot, (2) absence of terminal discs on fingers and toes, (3) absence of circumarginal grooves on the fingers and toes, and (4) absence of tarsal folds. Given their phylogenetic and morphological distinctiveness, we assign them to the oldest available generic name for this clade, Liurana Dubois 1987, and transfer Liurana from Dicroglossidae to the family Ceratobatrachidae. In contrast, Ingerana tasanae was found to be clustered with strong support with the recently described genus Alcalus (Ceratobatrachidae), a small clade of otherwise Sundaic species; this constitutes a new record of the family Ceratobatrachidae for Myanmar and Thailand. Finally, Ingerana borealis clustered with the "true" Ingerana (family Dicroglossidae), for which the type species is I. tenasserimensis.

Advances in herpetological research emanating from China.

Advances in herpetological research emanating from China.

Cuticular features of Cryptocarya (Lauraceae) from Peninsular Malaysia, Thailand and Indo-China and its taxonomic implications

Cuticular features potentially offer valuable information on phytotaxonomy, especially for plants that are difficult to classify or identify. Cryptocarya is one such difficult and poorly known genus, and we investigated its cuticular features to evaluate their taxonomic implications. We examined cutinized leaf epidermis and the stomatal complex for 21 species from Peninsular Malaysia, Thailand and Indo-China, as well as one species endemic to Borneo and one to Sumatra, and compared features among members of this group, between this group and congeners in the other countries, and between this group and the other genera of Lauraceae. Many of the Cryptocarya species studied have straight to slightly curved anticlinal epidermal walls and butterfly-shaped stomatal ledges, although some variation was seen in the ornamentation of the periclinal epidermal walls and the appearance of the stomatal complex. Based on these results, we recognized four groups and two subgroups among the South-east Asian species. Comparison with congeneric taxa in the other countries indicates that butterfly-shaped stomatal ledges are often shared among the species across the regions. However, no cuticular features occur exclusively in any of the species groups of Cryptocarya studied here, nor in the genus as a whole. We consider that cuticular features are useful in the recognition of infrageneric groups within Lauraceae, bearing in mind that they might have evolved in parallel in different genera.

Observations on the biology of Afrotropical Hesperiidae (Lepidoptera). Part 9. Hesperiinae incertae sedis: Zingiberales feeders, genera of unknown biology and an overview of the Hesperiinae incertae sedis.

The Afrotropical genera that have been recorded to feed on Zingiberales are documented. Partial life histories are presented for Erionota torus Evans (a South-East Asian species established in Mauritius), Semalea arela (Mabille), S. pulvina (Plötz), Xanthodisca vibius (Hewitson), X. rega (Mabille), Hypoleucis ophiusa (Hewitson), Caenides dacena (Hewitson), Osmodes adon (Mabille), Gretna cylinda (Hewitson) and Moltena fiara (Butler). Additional notes from the literature are provided on the genera Leona and Rhabdomantis. Notes on natural enemies of E. torus and M. fiara are included. We find that the Zingiberaceae and Costaceae feeding genera, Semalea, Xanthodiscus, Hypoleucis and Caenides (part) are united by a C-shaped raised rim to the prothoracic spiracle of the pupa. The pupa of Osmodes adon indicates this genus may have no close affinities to other Afrotropical genera for which the life history is known. The pupa of G. cylinda is unlike any other that we have documented and may reflect that this is the only species which we have found to be formed on the open leaf under surface rather than in a shelter. The early stages of M. fiara indicate affinities with Zophopetes and related genera. The paper concludes with a brief comparative discussion of the early stages of the Afrotropical Hesperiinae incertae sedis as a whole. There appear to be useful characters to group species by the ova and pupae but less so by the caterpillars. Based on pupae alone, the Hesperiinae incertae sedis might be divided into nine groups.

Induction Of Shoot Buds Multiplication OfEusideroxylon ZwageriTesym. and Binned (Borneo Ironwood) By Using Nodal Explants

Eusideroxylon zwageriis a tree of the tropical rainforest which is belongs to a family of Lauraceae. This species is one of the hardest timber species in Southeast Asia and unfortunately endangered in some part of Southeast Asia. The objective of this study was to determine the optimal culture medium for the induction of shoot buds from nodal explants of E. zwageri. Different concentrations and combinations of BAP (1.0, 2.0, 3.0, 4.0, 5.0 and 6.0 mg/L) alone or either in combination with NAA (0.5 mg/L) or IBA (0.5 mg/L) were used in order to induce multiple shoot buds of this species. The nodal explants were collected from the healthy lateral branches of two to three years old of E. zwageri sapling. The finding in this multiple shoot buds study showed that MS medium which have been supplemented with 5.0 mg/L of BAP alone or in combination with either 0.5 mg/L of NAA or IBA had induced the highest mean number of shoots buds and mean number of leaves respectively. It can be recommended that studies on plantlet regeneration of this species should be conducted and field performance should be carried out on this regenerated species.

Assessment of faecal glucocorticoid metabolite excretion in captive female fishing cats (Prionailurus viverinus) in Thailand.

There is little information on the endocrinology of fishing cats (Prionailurus viverinus), an endangered species in Southeast Asia, especially that pertaining to adrenal function. This study characterized faecal glucocorticoid metabolites in female fishing cats housed at Chiang Mai Night Safari to investigate seasonal and age relationships in hormone patterns. Faecal samples were collected 3 days/week for 1 year from seven females ranging in age from 4.5 to 9.6 years. A corticosterone enzyme immunoassay was validated for fishing cats by showing increases (∼60%) in faecal glucocorticoid immunoactivity above pre-treatment baseline levels within 1-2 days after an adrenocorticotrophic hormone injection. Faecal glucocorticoid metabolite concentrations were not related to age (P > 0.05), but there was a seasonal effect, with concentrations being higher (P < 0.05) during the winter (1.54 ± 0.04 µg/g) and rainy season (1.43 ± 0.04 µg/g) compared with the summer (1.22 ± 0.05 µg/g). Significant relationships were found between faecal glucocorticoids and rainfall (positive) and day length (negative), but not a temperature-humidity index. This is the first study to assess adrenal steroidogenic activity in female fishing cats, and we found that glucocorticoid metabolite production was influenced by seasonal factors, but not by age. We conclude that weather patterns should be taken into consideration in future studies of glucocorticoid activity in this endangered species, especially those studies aimed at improving captive management to create self-sustaining and healthy populations.

Genetic differentiation in two widespread, open-forest bird species of Southeast Asia (Copsychus saularis and Megalaima haemacephala): Insights from ecological niche modeling

Abstract Ecological niche modeling has emerged as an useful tool in the investigation of the phylogeographic histories of species or communities in a region. The high biodiversity (oftentimes cryptic), and complex geography and geological history of Southeast Asia particularly call for multipronged approaches in phylogeographic investigations. Past studies have focused on taxa that are associated with lowland rainforests, which is the dominant natural vegetation type. Here, we combine published phylogenetic data, ecological niche modeling and paleo-climate models to reveal potential drivers of divergence in two open-forest bird species, the oriental magpie-robin Copsychus saularis and Coppersmith barbet Megalaima haemacephala. In spite of broad overlap in current distributions, there are subtle differences in their climatic niches, which result in different responses to past climatic changes. For C. saularis, both Last Glacial Maximum climate models indicated that the entire Sundaland was climatically suitable, while phylogenetic analyses found divergent eastern and western Sundaland lineages. We thus postulate that this genetic divergence was a result of past separations of coastal habitats into eastern and western portions due to the emergence of Sunda shelf as sea-level fell. The current separation of morphological subspecies in Borneo is maintained by low climatic suitability (high annual rainfall) in certain regions. The extirpation of M. haemacephala from Borneo and southern Malay Peninsula might have been driven by unsuitable conditions (high temperature seasonality) in central Sundaland and/or the lack of open woodlands. Our study shows that ecological niche modeling adds a powerful dimension to our attempt to understand lineage evolution in space.

Rapid detection of SNPs in candidate genes regulating the growth of orange-spotted grouper, Epinephelus coioides (Hamilton, 1822), using semiconductor sequencing

Orange-spotted grouper (Epinephelus coioides) is one of the most important marine food species in Southeast Asia and China and has been cultured for decades. In this study, we fully utilized the limited capacity of semiconductor sequencing, the high efficiency of long-range PCR for target enrichment and a non-indexed pooling strategy to screen single-nucleotide polymorphisms (SNPs) in a breeding population of orange-spotted grouper. Forty-one genomic DNA fragments, with a total length of approximately 180 kb, including 22 candidate genes that control growth, and from a DNA pool of 20 heaviest and 22 lightest individuals of the sampled population were successfully sequenced using an Ion Torrent Personal Genome Machine. 3 503 466 clear reads were produced with a length of 192 ± 56 bp, 86.8% of which were mapped to the reference with an average coverage depth of 2567-fold and physical coverage of 98.8%. Finally, 1623 high-quality SNPs were adopted. Compared with Sanger sequencing of three random common regions, the sensitivity and specificity of our approach were 39.4% and 100.0% respectively. A mutation located at the third position of the previously labelled start codon of growth hormone receptor type 1 invalidated the start codon. Furthermore, comparison of the frequencies of genotypes and alleles of this site between the two extreme groups, prediction of signal peptide and identification of conservative mRNA sequences suggested that the functional start codon is likely located at the position of another downstream in-frame ATG in the mutant. These detected SNP markers will provide important tools for the selective breeding of orange-spotted grouper.

First record of the genus Acropimpla Townes, 1960 (Hymenoptera:Ichneumonidae: Pimplinae) from Thailand, with descriptions of three new species.

Nine species of Acropimpla Townes, 1960 are recorded from Thailand with three species described as new: A. leoni sp.n., A. natae sp.n., A. patellalutea sp.n. Six species, A. aspera Gupta & Tikar, 1976, A. hapaliae (Rao, 1953), A. leucostoma (Cameron, 1907), A. nakula Gupta & Tikar, 1976, A. taishunensis Liu, He & Chen, 2010, and A. uchidai (Cushman, 1933) are recorded from Thailand for the first time. A key to the females of Acropimpla species in South East Asia is also provided.

First record ofNeosiphonia echinata(Rhodomelaceae, Rhodophyta) in the South Pacific: an introduced species in Southeast Asia

AbstractNeosiphonia echinata,a well-known western Atlantic species, is here reported for the first time from Indonesia and this extends its distribution into Southeast Asia.Neosiphonia echinatawas collected from Sulawesi, Indonesia as an epiphyte onKappaphycus alvareziiand growing on ropes in a culture farm. We also collected this species from Florida, USA, and then compared their morphology and molecular data.Neosiphonia echinatais identified by having erect main filaments arising from prostrate filaments, numerous rhizoids cutting off from the proximal end of pericentral cells by a cross wall, four pericentral cells throughout the thallus, ecorticate axes, very long and abundant trichoblasts more than twice forked, abundant adventitious laterals, procarps with four-celled carpogonial branches, and a spiral arrangement of tetrasporangia. Our phylogenetic analyses ofrbcL andcox1 indicate thatN. echinatafound in Indonesia was very closely related to material from the Western Atlantic Ocean.Neosiphonia echinatamay be added as an introduced species to the Indonesian marine flora. The shipping routes western Atlantic-Mediterranean-Indonesia and western Atlantic-Pacific Ocean-Indonesia might be considered as possible pathways of introduction as a consequence of ballast water and hull fouling.

Ultra-structure of blood cells and molecular characteristics of Haemoproteus sp. in Blyth’s hawk eagle

Haemoproteus was detected in the blood smear of an adult female Blyth’s hawk eagle which was submitted to the Kasetsart University Raptor Rehabilitation Unit. The morphology and ultra-structure of leukocytes and gamonts of Haemoproteus were described. The gamont’s cytoplasm contained scattered brown to black pigment granules with a mean of 12 granules per parasite. Monocytes played a major role in the elimination of the gamonts. According to the annotated checklist of the avian Haemoproteidae, the Haemoproteus in this eagle was closely related to Haemoproteus buteonis. The phylogeny of mitochondrial cytochrome b genes of Haemoproteus revealed 96.5 and 97.6 % similarity in identity with the Haemoproteus from two collared scops owls and seven barn owls (99.3 to 99.8 % similarity) from Thailand, and they were also similar to the Haemoproteus isolated from other reported avian species in Southeast Asia. The eagle was clinically healthy without any treatment. It was free from Haemoproteus by light microscopy and molecular technique 8 months later and was released in Krungching Waterfall, Khao Luang National Park, southern Thailand.

Comparative study on volatile oils of four Panax genus species in Southeast Asia by gas chromatography–mass spectrometry

Four Panax genus species (Panax notoginseng (PN), Panax stipuleanatus (PS), Panax vietnamensis (PV) and Panax japonicus (PJ)) are all distributed in Southeast Asia. Besides PN, the other three species attracted little attention because of their limited distribution area, less wide application or relatively low quantity. PN had been cultivated as an industrial crop for hundreds years in China while the other three species as substitutes for PN in minority ethnic groups had no large-scale cultivation until now. The saponins of the four medical plants had been investigated in previous study while there has been virtually no reports describing their other compounds. In this study, the gas chromatography-mass spectrometry (GC–MS) was used to compare the volatile oils compounds of the four species. A total of 146 compounds were identified among all the species and every species contained compounds with similar quantity (all about 70). The falcarinol (28.86%) and andrographolide (38.35%) were the main compositions for PN. Furthermore, falcarinol was the dominant composition for PV (70.65%) and PS (64.61%), while pentadecanoic acid and ledene oxide-(II) were the high content compounds for PJ, accounting 9.31% and 8.39%, respectively. In addition, hierarchical clustering analysis was conducted to evaluate the relationship between the four Panax species based on contents (%) of chemical constituents in volatile oils. The results indicated that the PV and PS exhibited high similarity of volatile oils compounds. The PN, PV and PS could be used as the raw materials of falcarinol presenting well biological activities.

Intraspecific variability of the facultative meiotic parthenogenetic root-knot nematode (Meloidogyne graminicola) from rice fields in Vietnam

Twenty years ago, the facultative meiotic parthenogenetic root-knot nematode (RKN), Meloidogyne graminicola, was recognised as an important rice pathogen in South Vietnam. Although this country is one of the most important rice exporters worldwide, a comprehensive picture of the occurrence of M. graminicola in Vietnamese rice fields is still not available. Therefore a nematode survey was carried out with the aim of better understanding the geographical distribution, and the pathogenic and genetic variability of the RKN in Vietnam. From the fields surveyed in a range of ecosystems, 21 RKN populations were recovered from infected rice roots. A diagnostic SCAR marker was developed showing that all Vietnamese populations belong to M. graminicola. Furthermore, sequencing of the Internal Transcribed Spacer (ITS) of the rDNA genes confirmed this identification. These populations were then characterised using morphometrics and pathogenicity tests (host plant range diversity, reproduction and virulence diversity) revealing intraspecific variability. We showed that morphometric traits are mainly genetically heritable characters with significant differences among the studied populations. Finally, a distinctive trait signature was found for the populations isolated from the upland rice cultures. All together, our study reveals the prevalence of M. graminicola populations in Vietnamese rice. Further investigations need to be developed to explore the population dynamics and evolutionary history of this species in South East Asia.

Foliose Halymenia species (Halymeniaceae, Rhodophyta) from Southeast Asia, including a new species, Halymenia malaysiana sp. nov.

Abstract Despite the large number of species discovered in Halymenia, many remain poorly known due to the scarce information available. In order to facilitate species discrimination of foliose Halymenia species in Southeast Asia, molecular analysis and morphological studies were made on Halymenia collections from Malaysia, the Philippines, and Indonesia. The rbcL phylogenetic analyses showed that there are at least six taxa of foliose Halymenia occurring in Southeast Asia. Among the six taxa, a new species, Halymenia malaysiana P.-L. Tan, P.-E. Lim, S.-M. Lin et S.-M. Phang, is proposed based on both rbcL sequence analyses and morphological observations. Halymenia malaysiana is characterized by thalli possessing oblong or suborbiculate blades with a supple cartilaginous structure and gelatinous (slimy) texture, arising from a small discoid holdfast without a stipe, abruptly expanding into a broad blade and having a smooth surface with sinusoidally undulated margins. The phylogenetic analyses also revealed that Halymenia is a polyphyletic genus, which requires further taxonomic studies.

Mixed Mating System Are Regulated by Fecundity in Shorea curtisii (Dipterocarpaceae) as Revealed by Comparison under Different Pollen Limited Conditions

The maintenance of mixed mating was studied in Shorea curtisii, a dominant and widely distributed dipterocarp species in Southeast Asia. Paternity and hierarchical Bayesian analyses were used to estimate the parameters of pollen dispersal kernel, male fecundity and self-pollen affinity. We hypothesized that partial self incompatibility and/or inbreeding depression reduce the number of selfed seeds if the mother trees receive sufficient pollen, whereas reproductive assurance increases the numbers of selfed seeds under low amounts of pollen. Comparison of estimated parameters of self-pollen affinity between high density undisturbed and low density selectively logged forests indicated that self-pollen was selectively excluded from mating in the former, probably due to partial self incompatibility or inbreeding depression until seed maturation. By estimating the self-pollen affinity of each mother tree in both forests, mother trees with higher amount of self-pollen indicated significance of self-pollen affinity with negative estimated value. The exclusion of self-fertilization and/or inbreeding depression during seed maturation occurred in the mother trees with large female fecundity, whereas reproductive assurance increased self-fertilization in the mother trees with lower female fecundity.

Bacterial community associated with the intestinal tract of Chinese mitten crab (Eriocheir sinensis) farmed in Lake Tai, China.

Chinese mitten crab (CMC, Eriocheir sinensis) is an economically valuable species in South-East Asia that has been widely farmed in China. Characterization of the intestinal bacterial diversity of CMC will provide insights into the aquaculturing of CMCs. Based on the analysis of cloned 16S rRNA genes from culture-independent CMC gut bacteria, 124 out of 128 different clones reveal ＞95% nucleotide similarity to the species belonging to the four phyla of Tenericutes, Bacteroidetes, Firmicutes and Proteobacteria; one clone shows 91% sequence similarity to the member of TM7 (a candidate phylum without cultured representatives). Fluorescent in situ hybridization also reveals the abundance of Bacteroidetes in crab intestine. Electron micrographs show that spherical and filamentous bacteria are closely associated with the microvillus brush border of the midgut epithelium and are often inserted into the space between the microvilli using a stalk-like cell appendage. In contrast, the predominant rod-shaped bacteria in the hindgut are tightly attached to the epithelium surface by an unusual pili-like structure. Both 16S rRNA gene denaturing gel gradient electrophoresis and metagenome library indicate that the CMC Mollicutes group 2 appears to be present in both the midgut and hindgut with no significant difference in abundance. The CMC Mollicutes group 1, however, was found mostly in the midgut of CMCs. The CMC gut Mollicutes phylotypes appear to be most closely related to Mollicutes symbionts detected in the gut of isopods (Crustacea: Isopoda). Overall, the results suggest that CMCs harbor diverse, novel and specific gut bacteria, which are likely to live in close relationships with the CMC host.

Two new synonyms among Chinese Thripinae (Thysanoptera: Thripidae).

Two species of Thripinae described from China, Danothrips dianellae Zhang and Tong, and Taeniothrips calopgomi Zhang, are here recognized as synonyms of two widespread Southeast Asian species.