New earthworm species of the genus Amynthas Kinberg, 1867 from Thailand (Clitellata, Oligochaeta, Megascolecidae)

Four new species of terrestrial earthworms from the zebrus-group in the genus Amynthas Kinberg, 1867, are described from Nan province, north Thailand: Amynthas phatubensissp. n., from Tham Pha Tub Arboretum, Amynthas tontongsp. n., from Tontong Waterfall, Amynthas borealissp. n., from Chaloemprakiat district, and Amynthas srinansp. n., from Srinan National Park.After comparing with the two closely related Laos species Amynthas chandyi Hong, 2008 and Amynthas namphouinensis Hong, 2008, the four new species show clear morphological differences, and also it is confirmed that there are no previous records of the species described here. Amynthas phatubensissp. n. is the largest (longest) sized of these earthworms and is the only species that lives in limestone habitats. The genital characters are different among them and also from the two Laotian species. Molecular systematics would be a good method for further analysis of the diversity and species boundaries in SE Asian Amynthas.     

Megalohypha, a new genus in the Jahnulales from aquatic habitats in the tropics

A new fungus collected from submerged wood in tropical forest streams in Panama and Thailand is described as a new genus and species in the Jahnulales. This fungus, Megalohypha aqua-dulces, is described based on ascospore morphology, which differs substantially from that of the other genera in the Jahnulales. It has these morphological features: ascomata hyaline, translucent, with subtending, wide, brown, septate, stoloniferous hyphae; peridium of large, thin-walled cells; hamathecium of septate pseudoparaphyses; asci clavate, fissitunicate, 8-spored; and ascospores 1-septate, brown, rough-walled, with longitudinal sulcate striations. An interesting feature of Megalohypha aqua-dulces, which it shares with some other taxa in Jahnulales, is the presence of both sessile and stalked fruiting bodies. In addition, the stalks have morphology that consists of a wide hypha with a narrower hypha apparently enclosed within and the stalks are strongly constricted at the septa.     

Integrative taxonomy reveals phenotypic plasticity in the freshwater mussel Contradens contradens (Bivalvia: Unionidae) in Thailand,with a description of a new species

The freshwater mussel Contradens contradens (Lea, 1838) occurs in most types of freshwater habitats throughout Thailand. The species shows extensive variation in shell morphology, which has led to the recognition of six different subspecies. In this study, the validity of these six subspecies plus one unknown species was assessed using an integrative taxonomic approach. Geometric morphometric analyses revealed significant differences in shell shapes among these six nominal morphological subspecies, although a considerable degree of overlap was detected in some groups. In contrast, the phylogenetic tree obtained from the concatenated data of mitochondrial COI and nuclear H3 gene sequences and molecular species delimitation analyses revealed only three supported clades. These clades are proposed herein as three distinct species, and strongly corresponded to the biogeographically disjunct drainage systems in Thailand. They consisted of the (i) C. contradens clade found in the Chao Phraya Basin and other rivers that drain into the Gulf of Thailand, (ii) C. crossei clade that is restricted to the Middle Mekong Basin, and (iii) a clade containing only the newly discovered species from Huai Luang River in the north-east of Thailand, which is described herein as Contradens rolfbrandti Jeratthitikul & Panha, sp. nov. Speciation among these congeners was probably caused by the restriction of gene flow due to the past geomorphology of the river systems. The intraspecific variation in the shell shape detected here does not reflect the evolution of the mussel, but rather is evidence of phenotypic plasticity.http://zoobank.org/urn:lsid:zoobank.org:pub:649B6093-E1DD-4FD8-8185-A4696C43AD36; http://zoobank.org/urn:lsid:zoobank.org:act:777DDE86-8397-4FF3-AA78-0BE0F34611F4     

New haplotype of the complete mitochondrial genome of <Emphasis Type="Italic">Crocodylus siamensis</Emphasis> and its species-specific DNA markers: distinguishing <Emphasis Type="Italic">C. siamensis</Emphasis> from <Emphasis Type="Italic">C. porosus</Emphasis> in Thailand

Based on molecular phylogeny of available complete mitochondrial DNA (mtDNA) genome sequences reveals that Crocodylus siamensis and C. porosus are closely related species. Yet, the sequence divergence of their mtDNA showed only a few values under conspecific level. In this study, a new haplotype (haplotype2, EF581859) of the complete mtDNA genome of Siamese crocodile (C. siamensis) was determined. The genome organization, which appeared to be highly similar to haplotype1 (DQ353946) mtDNA genome of C. siamensis, was 16,814 bp in length. However, the sequence divergence between the two genomes differed by around 7–10 and 0.7–2.1% for the haplotype1 between C. siamensis and C. porosus (AJ810453). These results were consistent with the phylogenetic relationship among the three genomes, suggesting that C. siamensis haplotype1 mtDNA genome might be the hybrid or the intraspecific variation of C. porosus. On the other hand, our specimen was found to be a true C. siamensis. Simultaneously, the seven species-specific DNA markers designed based on the distinctive site between haplotype2 mtDNA sequences of C. siamensis and haplotype1 mtDNA sequence of C. siamensis–C. porosus were successfully used to distinguish C. siamensis from C. porosus. These effective markers could be used primarily for rapid and accurate species identification in population, ecology and conservation studies.     

Isolation of ergosterol peroxide from Nomuraea rileyi infected larvae of tobacco cutworm

In the search for potential cytotoxic substances produced by Nomuraea rileyi, an active compound was isolated from mycosed insects through an activity guided fractionation process. The compound, cytotoxic against the Sf9 insect cell line, was identified to be ergosterol peroxide (5α, 8α-epidioxy-24(R)-methylcholesta-6, 22-dien-3β-ol) using nuclear magnetic resonance techniques, infrared spectrometry, and mass spectroscopy. Anticancer screens demonstrated that ergosterol peroxide at micromolar concentrations inhibited the growth of hormone-dependent breast cancer cell line (T47D), hormone-independent breast cancer cell line (MDA-MB-231), human epidermoid carcinoma in mouth cell line (KB), human cervical carcinoma cell line (HeLa), lung cancer cell line (H69AR) and human cholangiocarcinoma cell line (HuCCA-1). Ergosterol peroxide showed moderate effects against Spodoptera litura larvae; 46.7% mortality via topical application after 7 day post-treatment whereas the insect’s death was not found in per os application. The amounts of ergosterol peroxide produced by N. rileyi cultures under in vitro and in vivo were determined. The physiological levels of ergosterol peroxide detected in mycosed and mummified cadavers were very low (0.011 and 0.386 μg/larva) less then levels that either inhibited insect cell proliferation or caused insecticidal activity.     

Automated calculation of docking of artemisinin to heme

We report automated molecular docking of artemisinin to heme. The effects of atomic charges, and ligand and heme structures on the docking results were investigated. Several charge schemes for both artemisinin and heme, artemisinin structures taken from various optimization methods and X-ray data, and five heme models, were employed for this purpose. The docking showed that artemisinin approaches heme by pointing O1 at the endoperoxide linkage toward the iron center, a mechanism that is controlled by steric hindrance. This result differs from that reported by Shukla et al. which suggested that heme binds with artemisinin at the O2 position. The docking results also depended on the structures of both artemisinin and heme. Moreover, the atomic charges of heme have a significant effect on the docking configurations.     

The genusBrachydesmiella from submerged wood in the tropics, including a new species and a new combination

During studies of fungi occurring on wood submerged in freshwater streams in the tropics, five species ofBrachydesmiella, namely,B. anthostomelloidea, B. biseptata, B. caudata, B. orientalis comb. nov., andB. verrucosa sp. nov., were collected. There are described and illustrated from natural substrata, wooden test blocks, and from pure culture, based mainly on collections made in Thailand.Brachydesmiella verrucosa differs from the other four species in having 3-celled, coarsely verrucose, ampoule-shaped conidia, with a long, thin, cylindrical, verruculose apical cell. A key toBrachydesmiella species is provided.     

Citric acid production by 2-deoxyglucose-resistant mutant strains of Aspergillus niger

Summary Many mutant strains showing resistance to 2-deoxy-d-glucose (DG) on minimal medium containing glycerol as a carbon source were induced from Aspergillus niger WU-2223L, a citric acid-producing strain. The mutant strains were classifiable into two types according to their growth characteristics. On the agar plates containing glucose as a sole carbon source, mutant strains of the first type showed good growth irrespective of the presence or absence of DG. When cultivated in shake cultures, some strains of the first type, such as DGR1–2, showed faster glucose consumption and growth than strain WU-2223L. The period for citric acid production shortened from 9 days for strain WU-2223L to 6–7 days for these mutant strains. The levels and yields of citric acid production of the mutant strains were almost the same as those of strain WU-2223L. The mutant strains of the second type, however, showed very slow or no growth on both the agar plates containing glucose and fructose as sole carbon sources. In shake cultures, mutant strains such as DGR2-8 showed decreased glucose consumption rates, resulting in very low production of citric acid.