Genetic barcoding systematically comparing genes in del(5q) MDS reveals a central role for CSNK1A1 in clonal expansion

How genetic haploinsufficiency contributes to the clonal dominance of hematopoietic stem cells (HSC) in del(5q) myelodysplastic syndrome (MDS) remains unresolved. Using a genetic barcoding strategy, a systematic comparison was carried out on genes implicated in the pathogenesis of del(5q) MDS in direct competition with each other and wild-type (WT) cells with single clone resolution. Csnk1a1 haploinsufficient HSCs expanded (oligo)clonally and outcompeted all other tested genes and combinations. Csnk1a1-/+ multipotent progenitors showed a pro-proliferative gene signature and HSCs a downregulation of inflammatory signaling/immune response. In validation experiments, Csnk1a1-/+ HSCs outperformed their WT counterparts under a chronic inflammation stimulus, also known to be caused by neighboring genes on chromosome 5. A crucial role for Csnk1a1 haploinsufficiency in the selective advantage of the 5q- HSC is therefore proposed. It is implemented by creation of a unique competitive advantage through increased HSC self-renewal and proliferation capacity, as well as increased fitness under inflammatory stress.

The unusual genus Ramiheithrus (Trichoptera: Philorheithridae)

The descriptions of males of the two described species of Ramiheithrus (R. virgatus Neboiss and R. kocinus Neboiss) are expanded using additional material. The female, pupa and larva of Ramiheithrus virgatus are described for the first time for the genus. Preliminary genetic barcoding analysis suggests the presence of additional undescribed species that are only known as larvae. The unusual larval habitat is described and issues relating to this are discussed.

Integrative description of new giant pill-millipedes from southern Thailand (Diplopoda, Sphaerotheriida, Zephroniidae)

Two new species of giant pill-millipedes, Zephronia viridisoma Rosenmejer & Wesener sp. nov. and Sphaerobelum aesculus Rosenmejer & Wesener sp. nov., are described based on museum samples from southern Thailand. Zephronia viridisoma sp. nov. comes from Khao Lak, while the type locality of S. aesculus sp. nov. is on Phuket Island. Both species are described integratively, combining light microscopy, scanning electron microscopy, multi-layer photography, micro-CT scans and genetic barcoding. Genetic barcoding was successfully conducted for holotypes of both new species, which could be added to a dataset of all published sequences of the family Zephroniidae, including all described species from Thailand, Laos and Cambodia up to 2020. Genetic barcoding of the COI gene revealed another female of S. aesculus sp. nov., 160 km east of the type locality. Both new species are genetically distant from all other Zephroniidae from Thailand and surrounding countries, showing uncorrected p-distances of 16.8–23.1%. A virtual cybertype of a paratype of Z. viridisoma sp. nov. was created and made publically accessible.

GENETIC BARCODING AND BOTANICAL STUDIES ON POTHOS SCANDENS L. LEAVES-AN EXTRA-PHARMACOPOEIAL DRUG OF AYURVEDA

Objective: This study was undertaken to carry out genetic barcoding, pharmacognostical and phytochemical studies on leaves of Pothos scandens L. belonging to the family Araceae. Methods: P. scandens, was collected from Cochin, Kerala, during the month of August 2019. Its leaves were examined for pharmacognostical characters and physicochemical, qualitative analysis and HPTLC studies was carried out following standard protocols recommended by the Ayurvedic Pharmacopoeia of India. Genetic barcoding of the fresh tender leaves samples was carried out through following standard methods. Results: Leaves are simple, alternate, broadly winged petiole. Transverse section through winged petiole portion showed U-shaped, epidermal cell along with cluster crystals. Transverse section through midrib boat-shaped structure, mesophyll undifferentiated, consist parenchyma cell, epidermis with thick cuticle along with waxy coating. Leaves powder shows green in color and microscopy reveals the presence of prismatic crystals, raphides, fibres, acicular crystals and monocot type 1 stomata. The plant is identified as Pothos scandens based on sequence homology and phylogenic analysis. Qualitative results show the presence of carbohydrates, protein, alkaloids and tannins. HPTLC profile shows 7 spots at both short UV and long UV and ferulic acid and kaempferol were identified through quantitative HPTLC. Conclusion: Pothos scandens is a climber with simple, alternate leaves. Acicular crystals and raphides, mucilaginous cavities, undifferentiated mesophyll are key characters of the identification of its leaf. DNA Barcoding results helps in correct identification of Pothos scandens. The results obtained from the phytochemical study will be helpful in the determination of the strength of purity and also for further standardization of the plant.

Seven new giant pill-millipede species and numerous new records of the genus Zoosphaerium from Madagascar (Diplopoda, Sphaerotheriida, Arthrosphaeridae)

Seven new species of the giant pill-millipede genus Zoosphaerium Pocock, 1895 are described from Madagascar: Z. nigrum sp. nov., Z. silens sp. nov., Z. ambatovaky sp. nov., Z. beanka sp. nov., Z. voahangy sp. nov., Z. masoala sp. nov. and Z. spinopiligerum sp. nov. All species are described based on drawings and scanning electron microscopy, while genetic barcoding of the COI gene was successful for six of the seven new species. Additional COI barcode information is provided for the first time for Z. album Wesener, 2009 and Z. libidinosum (de Saussure & Zehntner, 1897). Zoosphaerium nigrum sp. nov. and Z. silens sp. nov. belong to the Z. libidinosum species-group, Z. ambatovaky sp. nov. to the Z. coquerelianum species-group, Z. beanka sp. nov., Z. voahangy sp. nov. and Z. masoala sp. nov. to the Z. platylabum species-group and Z. spinopiligerum sp. nov. to the Z. piligerum species-group. Updated identification keys are provided for each species-group. Two western dry forest species, Z. silens sp. nov. and Z. voahangy sp. nov. are recorded from two localities, while the other five species are currently only known from their type localities. Of special conservation concern might be Z. ambatovaky sp. nov. from the lowland rainforest fragment of Ambatovaky, a nowadays isolated lowland rainforest, rapidly shrinking due to slash and burn agriculture. In addition to the new species, new locality data is provided for 11 species and numerous unidentifiable species of Zoosphaerium: Z. neptunus (Butler, 1872), Z. platylabum (de Saussure & Zehntner, 1902) and Z. piligerum (de Saussure & Zehntner, 1897) from the central eastern montane forests, as well as Z. ambrense Wesener, 2009, Z. aureum Wesener, 2009, Z. libidinosum, Z. corystoides Wesener, 2009, Z. discolor Wesener, 2009, Z. smaragdinum Wesener, 2009, Z. villosum Wesener & Sierwald, 2005 and Microsphaerotherium anjozorobe Wesener, 2009.

A new species of Leptopelis (Anura, Arthroleptidae) from the south-eastern slope of the Ethiopian Highlands, with notes on the Leptopelis gramineus species complex and the revalidation of a previously synonymised species

A new ground-dwelling species of treefrog in the genus Leptopelis is described from the Harenna Forest in south-eastern Ethiopia. The description is based on morphology and acoustics and is supported by molecular data. The new species has a small body size, and the digital discs on fingers and toes are significantly more conspicuous than in other semi-fossorial members of the L. gramineus complex. It occupies forest habitats at lower altitudes and is separated ecologically and geographically from high-altitude species of the complex. One of them, a parapatric cryptic species from Bale and Arsi Mountains, is resurrected from synonymy of L. gramineus and given a new name, L. montanus. Genetic barcoding of specimens from both populations showed that they belong to two distinct lineages that had been revealed by recent phylogenetic research. To confirm the geographic separation of the studied populations, the collection area of L. gramineus types was verified through analysis of the diary and the final report of the 2nd expedition of V. Bottego, and through matching of the route described in it with modern maps. The type locality of L. gramineus sensu stricto is restricted to Gamo Gofa, Ethiopia. Following the results of recent phylogenetic studies, the range of L. gramineus is limited to west of the Great Rift Valley. An identification key to the named Ethiopian species of the genus is provided.

Genetic barcoding of museum eggshell improves data integrity of avian biological collections

Natural history collections are often plagued by missing or inaccurate metadata for collection items, particularly for specimens that are difficult to verify or rare. Avian eggshell in particular can be challenging to identify due to extensive morphological ambiguity among taxa. Species identifications can be improved using DNA extracted from museum eggshell; however, the suitability of current methods for use on small museum eggshell specimens has not been rigorously tested, hindering uptake. In this study, we compare three sampling methodologies to genetically identify 45 data-poor eggshell specimens, including a putatively extinct bird’s egg. Using an optimised drilling technique to retrieve eggshell powder, we demonstrate that sufficient DNA for molecular identification can be obtained from even the tiniest eggshells without significant alteration to the specimen’s appearance or integrity. This method proved superior to swabbing the external surface or sampling the interior; however, we also show that these methods can be viable alternatives. We then applied our drilling method to confirm that a purported clutch of Paradise Parrot eggs collected 40 years after the species’ accepted extinction date were falsely identified, laying to rest a 53-year-old ornithological controversy. Thus, even the smallest museum eggshells can offer new insights into old questions.

Another hitchhiker exposed: Diceratocephala boschmai (Platyhelminthes: Temnocephalida) found associated with ornamental crayfish Cherax spp.

There are known plenty tiny invertebrate species associated with decapod crustaceans. Our contribution documents the first record of New Guinean ornamental Cherax crayfish epibiont, Diceratocephala boschmai (Platyhelminthes: Temnocephalida) found in pet trade in the Czech Republic. Correct species assignment was confirmed by genetic barcoding. The pathway for introductions of organisms unintentionally moved in association with ornamental crayfish via the international trade has been highlighted. Although Cherax species survive in thermal waters in Europe, even potential introduction to these habitats posed threat to native crayfish ectosymbionts (branchiobdellids) thought similar ecological niche.