oca2 targeting using CRISPR/Cas9 in the Malawi cichlid Astatotilapia calliptera

Identifying genetic loci underlying trait variation provides insights into the mechanisms of diversification, but demonstrating causality and characterising the role of genetic loci requires testing candidate gene function, often in non-model species. Here we establish CRISPR/Cas9 editing in Astatotilapia calliptera, a generalist cichlid of the remarkably diverse Lake Malawi radiation. By targeting the gene oca2 required for melanin synthesis in other vertebrate species, we show efficient editing and germline transmission. Gene edits include indels in the coding region, likely a result of non-homologous end joining, and a large deletion in the 3′ UTR due to homology-directed repair. We find that oca2 knock-out A. calliptera lack melanin, which may be useful for developmental imaging in embryos and studying colour pattern formation in adults. As A. calliptera resembles the presumed generalist ancestor of the Lake Malawi cichlids radiation, establishing genome editing in this species will facilitate investigating speciation, adaptation and trait diversification in this textbook radiation.

A New Species of Vietnamophryne with an Extended Description of Vietnamophryne orlovi

We describe Vietnamophryne cuongi sp. nov., a new species of microhylid frog from northern Vietnam, based on morphological and molecular differences. Although superficially similar to remaining Vietnamophryne members, the new species differs by a number of diagnostic morphological characters, such as body size, habitus, head dimensions, finger and toe morphology, skin texture, as well as colour pattern. Phylogenetic analyses based on a fragment of the mitochondrial 16S gene place the new species to be sister to V. orlovi, from which it differed by 2.4% genetic divergence. We also describe the first female known of V. orlovi, collected in the forest of Phia Oac-Phia Den National Park, Cao Bang Province. The latter species, for which we provide an extended description herein, was recently described based on the single male type specimen only from the same site in northern Vietnam. Vietnamophryne cuongi sp. nov. is only known from three specimens, two adult females and a juvenile, from the evergreen tropical forest of Ba Vi National Park, Hanoi. As such, it is likely to be at high risk of habitat loss. Considering its high ecological specialization and the small known distribution range of the new species, we propose Endangered as IUCN Red List status.

Image analysis of real-time classification of cherry fruit from colour features

An image analysis algorithm for the classification of cherries in real time by processing their digitalized colour images was developed, and tested. A set of five digitalized images of colour pattern, corresponding to five colour classes defined for commercial cherries, was characterized. The algorithm performs the segmentation of the cheery image by rejecting the pixels of the background and keeping the image features corresponding to the coloured area of the fruit. A histogram analysis was carried out for the RGB and HSV colour spaces, where the Red and Hue components showed differences between each of the specified colour patterns of the exporting reference system. This information led to the development of a hybrid Bayesian classification algorithm based on the components R and H. Its accuracy was tested with a set of cherry samples within the colour range of interest. The algorithm was implemented by means of a real time C++ code in Microsoft Visual Studio environment. When testing, the algorithm showed a 100% effectiveness in classifying a sample set of cherries into the five standardized cherry classes. The components of the hardware-software system for implementing the methodology are low cost, thus ensuring an affordable commercial deployment.

A new subspecies of Сryptocephalus ergenensis Morawitz, 1863 (Coleoptera, Chrysomelidae) from Kazakhstan and significance of colour pattern polymorphism examination

Cryptocephalus apicalis species group is introduced and defined within the subgenus Asionus Lopatin, 1988. Cryptocephalus ergenensis Morawitz, 1863 from this group is first recorded in Eastern Kazakhstan and this form with unusual colour pattern is described herein as a new subspecies kalbensis Mikhailov, ssp. nov. The members of Cryptocephalus apicalis species group are keyed and illustrated. Significance of the colour pattern polymorphism in taxonomy is discussed.

Convergent morphology and divergent phenology promote the coexistence of Morpho butterfly species

The coexistence of closely-related species in sympatry is puzzling because ecological niche proximity imposes strong competition and reproductive interference. A striking example is the widespread wing pattern convergence of several blue-banded Morpho butterfly species with overlapping ranges of distribution. Here we perform a series of field experiments using flying Morpho dummies placed in a natural habitat. We show that similarity in wing colour pattern indeed leads to interspecific territoriality and courtship among sympatric species. In spite of such behavioural interference, demographic inference from genomic data shows that sympatric closely-related Morpho species are genetically isolated. Mark-recapture experiments in the two most closely-related species unravel a strong temporal segregation in patrolling activity of males. Such divergence in phenology reduces the costs of reproductive interference while simultaneously preserving the benefits of convergence in non-reproductive traits in response to common ecological pressures. Henceforth, the evolution of multiple traits may favour species diversification in sympatry by partitioning niche in different dimensions.

Variations in phenotypic characteristics of Vigna unguiculata (L) Walp. seeds sourced from open markets in Benin City, Nigeria.

Vigna unguiculata, also known as a 'cowpea,' is a crop of numerous varieties. In Benin City, Nigeria, three common varieties include "Ife Brown", "Ekpoma Local", and "Sokoto White". Being a very common delicacy and perhaps a very cheap source of protein for the populous, it is sold in nearly all available open markets. The crop is mainly consigned from the numerous farms in the northern part of the country and thus vary amenable to genetic diversity. This study was undertaken to investigate whether there were any major differences within and between these varieties when sourced from different locations in selected markets in Benin City. The seeds were obtained from major open store markets in and around Benin City, Edo State, and were morphologically characterized quantitatively and qualitatively using standard descriptors. Seed length, width, thickness, weight, and volume were among the quantitative parameters, while the qualitative parameters included seed brilliance, shape, eye colour, eye pattern, splitting of testa, testa texture, basal colour, pattern of variegation, colour of variegation, and basal colour of variegated seeds. There were no variations in the qualitative parameters among the seeds of "Ekpoma Local" and "Sokoto White" varieties; however those of the "Ife Brown" variety were significant variable in seed colour. Significant variations existed in the seed quantitative parameters. Among seed parameters measured, seed volume was more diverse, with a coefficient of variation of 13.15 - 14.14, when compared with coefficients of variation for other quantitative parameters measured. In terms of seed volume therefore, the "Sokoto White" variety appeared to be more diverse than "Ekpoma Local" and then "Ife Brown" in that order. In terms of overall variation, group mean sum of squares for the "Ife Brown" variety was 146.95, compared to 26.18 and 31.23 for "Ekpoma Local" and "Sokoto White" respectively, indicating that the "Ife Brown" variety was the most likely variable species among the three in the Markets of Benin City.

Five new species of the sisorid catfish genus Glyptothorax from Iran (Teleostei: Sisoridae)

Five new species of Glyptothorax are described from Iran. Glyptothorax alidaeii, new species, from the Seimare in the Karkheh drainage, G. galaxias, new species, from the upper Karun drainage, G. hosseinpanahii, new species, from the Zohreh drainage, G. pallens, new species, from the Sirvan drainage, and G. shapuri, new species, from Shapur in the Helleh drainage. Glyptothorax silviae from the Jarrahi drainage is re-diagnosed. All six species are morphologically distinguishable by the structure of the thoracic adhesive apparatus, as well as morphometric characters and details in the colour pattern. They form distinct mitochondrial clades between 1.2% and 4.1% minimum K2P distance based on the mitochondrial DNA barcode region.

The term ‘marble’ in eighteenth-century encyclopaedic literature: from colourful and exclusive to grainy and popular

This article reports on findings concerning the use and understanding of marble in the eighteenth century, as uncovered by a team of geologists and conservators at the Department of Engineering Technology at the Technical University of Munich. While researching a group of marble objects in Bayreuth in order to devise suitable conservation methods, it became apparent that the eighteenth-century understanding of ‘marble’ was different to how we define the stone today. This earlier definition of marble was based on colour, pattern and the ability to shine when polished. However, by the end of the eighteenth century, there was a shift to a focus on the different grain sizes of the stones, while the previously defining quality of colour became less important. Such developments advanced towards the recognition of limestone and marble as two different types, enabling the distinction between sedimentary limestone and its metamorphic product marble to be drawn in the first half of the nineteenth century. At the same time, the exploration of local sources caused the exclusivity of marble to dwindle. Once a building and decorative material for the elite, it now became more widely available. Marble was still the material of sovereigns - proudly presented as locally found - but it simultaneously became accessible to a wider market for household utensils or collectors’ items. This is demonstrated through the exploration of a range of German sources, including encyclopaedias and lexicons with their inherent aim of accumulating the universal knowledge of their time, a ‘marble’ compendium, and a description of the prison and workhouse in St Georgen in Bayreuth, which had marble works on its premises.

Morphology and molecular phylogeny of ornamental freshwater prawns of the genus Macrobrachium (Decapoda, Caridea, Palaemonidae) from China with the description of a new species

This study dealt with three species of ornamental palaemonid freshwater prawns of the genus Macrobrachium, based on morphological and molecular analysis. Macrobrachium pentazona He, Gao & Guo, 2009; M. laevis Zheng, Chen & Guo, 2019; and M. bilineare sp. nov. are distinguishable from closely related species by segmental ratios, spination of the second pereiopods and the slender scaphocerite. Macrobrachium bilineare sp. nov. can easily be recognized in the field by its bright colour pattern. Molecular evidence of mitochondrial cytochrome oxidase subunit I (COI), also supports the characterization of this new species, raising the total number of Macrobrachium spp. known from China to 41. Detailed description, illustrations, colour photographs, habitat information, distribution maps and features of conservation significance are also briefly discussed.

Annotated and updated checklist of marine crabs (Decapoda: Brachyura) of Mozambique supported by morphological and molecular data from shelf and slope species of the “MOZAMBIQUE” surveys

An updated checklist of Mozambican marine brachyuran crabs is generated based on an exhaustive revision of the existing literature, together with the additional records provided by the specimens collected throughout the three “MOZAMBIQUE” surveys carried out in Mozambican waters during three consecutive years (2007–2009) by the Instituto Español de Oceanografía, (Spanish Institute of Oceanography, IEO). A total of 269 species, grouped in 15 superfamilies, 26 families and 172 genera are reported in the checklist, and a detailed inventory is produced with the list and remarks about the brachyuran species collected. Thirty-nine crab species belonging to 19 families were identified based on morphological characteristics and/or genetic tools. DNA barcode sequences (16S rRNA and/or COI) were obtained for 37 species, including 16S and COI sequences that are new for 26 and 14 species, respectively. Colour photographs of fresh specimens illustrate the comments about most species, being the first time that the original colour pattern is described for some of them. New records in Mozambican waters are reported for the species Paromolopsis boasi, Mursia aspera, Carcinoplax ischurodous, Tanaoa pustulosus, Euclosiana exquisita, Oxypleurodon difficilis, Naxioides robillardi, Samadinia galathea, Cyrtomaia gaillardi, Paramaja gibba, Pleistacantha ori, Parathranites granosus, Parathranites orientalis, Ovalipes iridescens and Charybdis smithii, and second records for Moloha alcocki, Samadinia pulchra and Charybdis africana. In addition, Raninoides crosnieri, S. galathea and P. ori were collected for the first time after their descriptions. The female of Samadinia galathea is described for the first time, and a potential new species of Mursia is reported. Some records expand the known bathymetric range of certain species and/or their general distribution. New molecular and morphological data suggest the necessity of the revision of P. boasi, R. crosnieri, C. africana and the genera Platymaia and Carcinoplax. The variability and taxonomic validity of some morphological characters in brachyuran systematic is discussed.