External Morphological Characters Research Articles

The new genus Purpurata (Lepidoptera, Crambidae, Spilomelinae), with descriptions of two new species from China

The male genitalia characters of four species, Botys iopasalis Walker, 1859, Pleuroptya obfuscalis Yamanaka, 1998, Botys plagiatalis Walker, 1859 and Patania shompen Singh et Ahmad, 2022, placed in the genus Patania Moore, 1888 before the present study, do not conform to the diagnosis of Patania. A new genus, Purpuratagen. nov., is established for these four species, and two new species, Purpurata directasp. nov. and Purpurata luridasp. nov. are described based on their external morphology and genitalia characters. Purpurata directasp. nov. is designated as the type species of the new genus. Five species of the new genus were clearly separated from Patania species in the Maximum likelihood phylogenetic tree constructed based on COI sequence data. Compared to Patania, the new genus Purpurata exhibits distinctive characters in male genitalia: the uncus is short, broad, and arc-shaped posteriorly; the gnathos is present and setose, or reduced; and the fibula is very small and setose. In addition, Patania clava (Xu & Du), syn. nov. is synonymized with Purpurata iopasaliscomb. nov. An identification key to species of the new genus is presented based on morphological characters of habitus and genitalia. Images of the habitus and genitalia are provided.

A new species of the genus Liotyphlops Peters, 1881 (Serpentes: Anomalepididae) from the Caribbean coast of Honduras

We describe a new species of the genus Liotyphlops from the Caribbean coast of Honduras in Nuclear Mesoamerica, a genus previously known only from Isthmian Mesoamerica and northern, central, and eastern South America. Our description is based on a single specimen found by chance. The new species differs from other congeners by unique combination of external morphological characteristics, specifically by the presence of three scales on the first vertical row of the dorsals, three supralabial scales, 425 dorsal scales, and dorsals in 22 series of uniformly pigmented scales across the body. The type locality represents the northernmost record for the genus and is located 920 km (airline) northwest of the previous record for Liotyphlops albirostris in Costa Rica. We developed an identification key for the genus and discussed natural history and biogeographical aspects of Liotyphlops, including the species described here.

Revealing the larval anatomy of the hoverfly Sphaerophoria rueppellii (Wiedemann, 1820) (Diptera, Syrphidae) using micro-computed tomography

Micro-computed tomography (micro-CT) is an X-ray-based technique that allows visualisation of the internal anatomy of insects in situ and does not require dissections. Traditionally, the study of insect anatomy has been mainly based on dissection techniques and microtome sections. However, micro-CT is becoming an increasingly widespread study technique. We report the use of micro-CT scans to study, in detail, the external and internal structures and organs of the third instar larva of the dipteran Syrphidae Sphaerophoria rueppellii, a Palaearctic species included in the list of effective agents used in European biocontrol programmes against aphids. Detailed images and videos provided evidence of the external morphological characteristics and an overview of the internal anatomy (musculature, digestive tube, salivary glands, Malpighian tubules, fat body, circulatory system, nervous system, cephalopharyngeal complex, tentorium, and mouth parts). The study evidenced the following: an external lateroventral complex of muscles that form muscular rings that allow the compression of the body segments in a similar functional way as occurs in annelid worms; the existence of mandibles with prosthecae; the nervous system present a conspicuous distinguishable suboesophageal ganglion; the so-called antennomaxillary organs are the antennae; the maxillae are separate structures, with an articulated maxillary palpus; S. rueppellii has only one pair of Malpighian tubules instead of the two pairs that were thought to be the general rule for Syrphid larvae; and the evidence of an imaginal disc of genitalia in the posterior part of the body, below the rectum, which in male larvae has a clear correspondence with the morphology of the external male genitalia, which allows to differentiate the sexes in the larval stage. This study constitutes a true anatomical atlas of the third instar larvae of S. rueppellii. High-quality rendered images and additional supplementary videos together with a 3D model, suitable for use with mobile devices, are useful tools for future research and teaching aids.

Expression of distal limb patterning genes in Hypsibius exemplaris indicate regionalization and suggest distal identity of tardigrade legs

BackgroundPanarthropods, a major group of invertebrate animals comprised of arthropods, onychophorans, and tardigrades, are the only limb-bearing members of Ecdysozoa. The complexity and versatility of panarthropod paired limbs has prompted great interest in their development to better understand the formation of these structures and the genes involved in this process. However, studies of limb patterning and development are overwhelmingly focused on arthropods, followed by select work on onychophorans but almost entirely lacking for tardigrades. This model organism bias is inherently limited and precludes a comparative analysis of how panarthropod legs originated, have evolved, and the likely limb patterning genes present in the earliest panarthropod ancestors. In this study, we investigated tardigrade homologs of seven arthropod distal limb patterning genes (apterous, aristaless, BarH1, clawless, Lim1, rotund, and spineless) to better characterize tardigrade limb development in a comparative context.ResultsWe detected homologs of all seven genes in the eutardigrade Hypsibius exemplaris and heterotardigrade Echiniscoides cf. sigismundi suggesting their conservation in both tardigrade lineages. Hybridization chain reaction experiments in H. exemplaris reveal a regionalized expression pattern for the genes aristaless, BarH1, clawless, rotund and spineless.ConclusionThe observed regionalized expression of the distal limb patterning genes in H. exemplaris might reflect the external morphological features of tardigrade legs, such as the distal claws, sensory organs in the proximal region, and specific muscle attachment sites. The comparison between the expression of these limb patterning genes in H. exemplaris relative to other panarthropods suggests their conserved role in the last common panarthropod ancestor, such as establishing the distal limb end and the distribution of sensory structures. Our results support the hypothesis that tardigrade legs are homologous to the distal region of other panarthropod limbs, as suggested by previous work on the expression of leg gap genes in H. exemplaris.

Cryodehydration applied to bovine fetal hearts: A model for studying cardiac organogenesis.

The study aimed to standardize the cryodehydration technique for bovine fetal hearts, focusing on optimizing protocols for each developmental stage to preserve morphological characteristics. We analyzed 29 bovine fetal hearts categorized into early, middle, and late developmental stages. These hearts underwent cryodehydration until a 60%-70% reduction in original fluid volume was achieved. Biometric data were recorded and statistically analyzed using Pearson correlation tests for age versus weight and age versus number of cryodehydration sessions. Morphometric comparisons before and after cryodehydration were performed using paired t-tests. In Group I, hearts exhibited well-defined structures, including the atrium cordis, ventriculus cordis, auricula atrii, aorta, truncus pulmonalis, and ramus coronaries arteria, which were preserved in Groups II and III. Additionally, in Group I the heart had a conical or flat apex cordis, whereas those in Groups II and III had a more pronounced apex. The average number of cryodehydration sessions required was 9.38 (±1.2) days for Group I, 12.37 (±1.4) days for Group II, and 15 days for Group III. A positive correlation was found between age and sample weight, indicating that more developed hearts were heavier. Similarly, there was a positive correlation between gestational age and the number of cryodehydration sessions, suggesting that more advanced stages required more cryodehydration sessions. Paired t-tests demonstrated high statistical significance in the morphometric parameters before and after cryodehydration, indicating a loss of mass during dehydration. However, there was no alteration in the macroscopic structure of the hearts, which remained morphologically preserved. In conclusion, cryodehydration shows promise for preserving and analyzing the external morphological characteristics of bovine fetal cardiac development. It also provides lightweight, odorless, and easy-to-handle specimens ideal for detailed morphological studies and offers a unique perspective for investigating cardiac morphology in biological research contexts.

Root and Canal Morphology of Permanent Maxillary and Mandibular Canines in a Sample of Yemeni Population Using Cone Beam Computed Tomography

Aims and Objective: This study aimed to investigate the external and internal morphological characteristics, including crown shape, root curvature, canal diameter, and root canal configuration, of maxillary and mandibular canine teeth in a Yemeni population using cone-beam computedtomography (CBCT). Materials and Methods: A total of 400 CBCT images (200 maxillary, 200 mandibular) were analyzed from 400 Yemeni subjects (averageage 34). Results: All maxillary canineshad a single root, while 4% of mandibular canineshad two roots (slightly higherin females, but not significantly). Maxillary canines were markedly larger in males (p = 0.001). Most maxillary canines(98.5%) had a single root canal, while 16.5% of mandibular canines had two (significantly higher in males). Using the Vertucci classification, root canal configuration analysis showed that maxillary canineswere predominantly Type I, while mandibular canines displayed more variation. Conclusion: This study provides insights into the morphological characteristics of Yemeni canine teeth. Gender is a significant factor in canine dimensions, with males having larger teeth.

Transcriptomic Communication between Nucleus and Mitochondria during the Browning Process of Lentinula edodes.

To explore the reason for cytoplasmic replacement's significant effect on browning, transcriptomic data of nuclear (N) and mitochondrial (M) mRNAs and long noncoding RNAs (lncRNAs) in L808 and two cytoplasmic hybrids (cybrids) (L808-A2 and L808-B) of Lentinula edodes at three different culturing times (80, 100, and 120 days) were obtained. The results showed that the expression of N and M genes and lncRNAs changed with the culture time and cytoplasmic source. Cytoplasmic replacement significantly affected some M and N genes related to the internal mechanism and external morphological characteristics of L. edodes browning. The internal browning mechanism should be the nicotinamide adenine dinucleotide phosphate (NADPH)-mediated antioxidant machinery to protect mycelia against oxidative stress induced by the generation of reactive oxygen species under light irradiation. External morphological characteristics were the changing features of brown films by melanin (an antioxidant) aggregation on the surface of the mycelia of the bag or log. Especially, some genes were related to the remodeling of the plasma membrane, extracellular enzymes of celluloses and hemicellulases, small molecules, and NADPH metabolic processes. Additionally, communication between the nucleus and mitochondria mediated by M-rps3 was reported for the first time, and it is mainly appreciated in M structural assembly, functional implementation, and cooperation with other organelles.

Integrative taxonomic review of the genus Rhodostrophia Hübner, 1823 and its allied genus Tanaotrichia Warren, 1893 (Lepidoptera: Geometridae) from the Western Himalaya

The genus Rhodostrophia Hübner, 1823 and its close ally Tanaotrichia Warren, 1893 (Geometridae: Sterrhinae) are reviewed from the North Western and Trans Himalayan regions of the Indian Himalaya along with some specimens from the Pakistan. In total, eight species were studied—seven Rhodostrophia spp. and one species of Tanaotrichia—with detailed morphological and genital characteristics. Rhodostrophia borealis stat. n. is raised from subspecies of R. cinerascens to species level after studying external morphology and distinct genitalia-based diagnostic features. However, this warrants further validation following the barcode sequencing of R. cinerascens. Illustrations of habitus, including type specimens, are provided along with images of genitalia and the sequence data of the mitochondrial cytochrome c oxidase subunit I (COI 5′ terminus) gene. A checklist of Indian Rhodostrophia and Tanaotrichia species is included with information on their documented distributional range.

Identification of morphologically similar genera based on reproductive systems and immature stages: a case study of Anastatus and Mesocomys (Hymenoptera: Eupelmidae)

Abstract Parallel or convergent evolution may result in phenotypically closely similar species. Many studies have reported that using solely traditional external morphological features can result in misidentification. Here, we discuss using the reproductive systems and immature developmental stages for scientific classification in Eupelmidae (Hymenoptera: Chalcidoidea), exemplifying with Anastatus Motschulsky and Mesocomys Cameron (Eupelmidae, Eupelminae). Results show that there are at least 6 distinct differences in the immature stages and the reproductive systems of the adults between species of the 2 genera: (1) the terminal ends of the left and right ovarioles are connected together in the studied species of Anastatus but not in Mesocomys, (2) the base of the lateral oviduct of Anastatus has a diverticulum that is lobate-shaped, whereas this diverticulum is spherical in Mesocomys, (3) the peduncle of the egg of Anastatus is significantly longer than that of Mesocomys, (4) the head capsule of the terminal instar larvae of Anastatus is uniformly translucent, whereas in Mesocomys it has a brown pattern, (5) the ocelli of the Anastatus pupa are smooth and slightly raised whereas the ocelli of Mesocomys each have a long filament each, and (6) the phallobase of adult male Anastatus is sharp basally but rounded in Mesocomys. Our findings not only provide novel insights for the differentiation of similar genera that can be difficult to distinguish in the immature stages but also provide a more comprehensive theoretical basis for clarifying the evolutionary and phylogenic status of Anastatus and Mesocomys.

Identification of Individuals of Two Takin Subspecies Using Biological and Ecological Criteria in Eastern Himalayas of China.

Limited background data are available on the Mishmi takin (Budorcas taxicolor taxicolor) and Bhutan takin (Budorcas taxicolor whitei) subspecies in the Eastern Himalayas of China because of the lack of systematic field investigations and research. Therefore, mature-animal ecological methods were used to evaluate these takin subspecies' phenotypic characteristics, distribution range, activity rhythm, and population size. From 2013 to 2022, 214 camera traps were installed for wild ungulate monitoring and investigation in all human-accessible areas of the Eastern Himalayas, resulting in 4837 distinguishable takin photographs. The external morphological characteristics were described and compared using visual data. Artificial image correction and related technologies were used to establish physical image models based on the differences between subspecies. MaxEnt niche and random encounter models obtained distribution ranges and population densities. Mishmi takins have a distribution area of 17,314 km2, population density of 0.1729 ± 0.0134 takins/km2, and population size of 2995 ± 232. Bhutan takins have a distribution area of 25,006 km2, population density of 0.1359 ± 0.0264 takins/km2, and population size of 3398 ± 660. Long-term monitoring data confirmed that the vertical migration within the mountain ecosystems is influenced by climate. Mishmi takins are active at 500-4500 m, whereas Bhutan takins are active at 1500-4500 m. The two subspecies were active at >3500 m from May to October yearly (rainy season). In addition, surveying combined with model simulation shows that the Yarlung Zangbo River is not an obstacle to migration. This study provides basic data that contribute to animal diversity knowledge in biodiversity hotspots of the Eastern Himalayas and detailed information and references for species identification, distribution range, and population characteristics.

Identification of body coloration related genes in ricefield eel (Monopterus albus) using GWAS

Body color is one of the most diverse external morphological characteristics of fish. It not only affects the function of fish predation and individual communication but also has important economic value. The ricefield eel (Monopterus albus) is a popular freshwater aquaculture fish species in China. Few studies have suggested some candidate genes involved in the formation of body color in this fish; however, further genetic evidences are required. Genome-wide association analysis (GWAS) is a powerful analytical method for identifying relationships between target traits and genetic markers or candidate genes in natural populations. In this study, two populations of ricefield eel with different body coloration were used to study the genetic factors that regulate the formation of body color. Through GWAS, 67 SNPs, 463 genes and 5 signaling pathways were found to be associated with body color. These results suggest that a complex molecular regulatory mechanism regulates ricefield eel body coloration. Our findings lay the foundation for subsequent genome selection breeding of ricefield eel.

Taxonomic Revision of the Dichotomius mormon (Ljungh, 1799) Species Group (Coleoptera: Scarabaeidae: Scarabaeinae), with a Description of a New Species from Bolivia.

Dichotomius Hope, 1838 (Coleoptera: Scarabaeidae: Scarabaeinae) is a very abundant and diverse genus of dung beetles of the New World ecosystems, with about 190 species, distributed in four subgenera. Luederwaldt (Separata Rev Mus Paul 14: 3-13 1929) proposed the division of subgenera into sections (now species groups) based mainly on characters of external morphology. Based on Luederwaldt's proposal, progress has been made in recent years in the taxonomic revision of the genus; however, inconsistencies have been found in the subgeneric division and species groups proposed by this author. Based on the external morphology and the male genital organ of the all type material and other material deposited in fifteen entomological collections, in this paper, the redefinition and taxonomic revision of the Dichotomius mormon species group is carried out. The new definition of the group and the species key are presented. Five species are included in the group: Dichotomius conicollis (Blanchard, 1846) (Bolivia), Dichotomius larseni sp. nov. (Bolivia), Dichotomius mormon (Ljungh, 1799) (Argentina, Brazil, and Paraguay), Dichotomius ohausi (Luederwaldt, 1923) (Brazil, Colombia, Ecuador, and Peru), and Dichotomius talaus (Erichson, 1847) stat. rev. (Bolivia, and Peru). The lectotypes for D. conicollis, Copris ephialtes Mannerheim, 1829, Copris nasuta Castelnau, 1840, Copris protensus Perty, 1830, D. ohausi, and D. talaus were designated. For each of the species that are included in the group, the following information is presented: taxonomic history, species' citation in published literature, description or redescription of males and females, list of material examined, photographs of the external morphology, illustrations of male genital organ and its endophallites, and distribution map.

A new threat to the European marine environment: The exotic polyclad flatworm Postenterogonia orbicularis (Schmarda, 1859)

Harbours and marinas are optimal places for the detection of the arrival of Non-Indigenous Species (NIS) in the early stages of introduction. Marine traffic serves as a route for species movement among different regions, while the species settlement in the ports is facilitated due to habitat degradation. Among NIS, polyclads flatworms are active hunters of bivalves, posing a threat to recipient environments and aquaculture. They can either use their pharynx and mucus to penetrate the mussels and oysters, or enter molluscs while they filter, making the management of the species difficult. In this regard, this work aims to report the first morphological and molecular identification of a new NIS polyclad native to the Pacific Ocean in Europe, specifically in the Ría de Avilés and the Gijón Port (North of Spain, Cantabrian Sea, Bay of Biscay). The occurrence of mature ovigerous specimens and the collection of specimens of various sizes from different times of the year, evidence a reproductively-active population in the area. The external morphological characters, as the ruffled pharynx and the cephalic eyes, and the internal ones, as the male copulatory apparatus, fit with the description of Postenterogonia orbicularis. The phylogenetic analysis based on the mitochondrial COI marker, placed the studied specimens from Avilés and Gijón (Spain) and the recently described species Idiostylochus tortuosus from Arcachon Bay (France) within the P. orbicularis clade (Ilyplanidae family). Therefore, our results suggest that I. tortuosus may be a synonym of the New Zealander P. orbicularis. Finally, we discuss the possible ecological impacts of its introduction and provide some management recommendations for controlling its influx in the recipient environments.

Whole-Genome Resequencing Reveals Signatures of Adaptive Evolution in Acanthopagrus latus and Rhabdosargus sarba.

Acanthopagrus latus and Rhabdosargus sarba are economically important marine species along the coast of China, with similar external morphological characteristics and living habits, with wide distribution and strong adaptability. To investigate the molecular mechanisms underlying the adaptive evolution of these two species, we conducted whole-genome resequencing of 10 individuals of both species from the coastal waters of Wuyu Island, Fujian, China, using high-throughput sequencing technology. We obtained SNP, InDel, CNV, and SV variation information and annotated these variations, constructing a genomic variation database for both species. By comparing the resequencing data with reference genomes, we identified 9,829,511 SNP loci in the population of A. latus and 34,051,056 SNP loci in the population of R. sarba. Using whole-genome SNP data, we employed Fst and ROD methods to identify candidate genomic regions under selection. Functional annotation and enrichment analysis using GO and KEGG databases revealed potential adaptive evolution in R. sarba associated with immune response, feeding, growth and development, and locomotion, while A. latus showed potential adaptive evolution associated with immune response, nervous system, growth and development, and metabolism.

The La Voulte-sur-Rhône Konservat-Lagerstätte reveals the male and female internal anatomy of the Middle Jurassic clawed lobster Eryma ventrosum

The biology of extinct animals is usually reconstructed from external morphological characters and comparison with present-day analogues. Internal soft organs are very rarely preserved in fossils and require high-tech approaches for visualization. Here, we report the internal anatomy of a female and male of the ~ 162 Myr-old lobster Eryma ventrosum from the Jurassic La Voulte-sur-Rhône Konservat-Lagerstätte in France using X-ray synchrotron tomography. The Erymidae is an extinct, species-rich, widespread and ecologically important Mesozoic family of decapod crustaceans. Our investigation revealed the anatomy of the locomotory, respiratory, circulatory, excretory, digestive, nervous and sensory, and reproductive systems at a resolution resembling low-magnification histology. Particularly notable is the detailed preservation of the small brain and the fragile hepatopancreas, the main metabolic organ of decapods that decays rapidly post-mortem. The remarkable preservation shows that the internal anatomy of Eryma ventrosum is closer to that of Nephropidae (clawed lobsters) than Astacidae (freshwater crayfish), their closest living relatives based on skeletal morphology. The microanatomy of the gonads and hepatopancreas indicates that the two specimens investigated were a young, well-nourished female and male prior to sexual maturity. The analysis of the soft anatomy reveals remarkable conservatism over 160 Myr and offers new insights into feeding, reproduction, life history and lifestyle of an important component of the macrozoobenthos of Middle Jurassic seas.

MORPHOTYPE MATTERS: AN EXPERIMENTAL ANALYSIS OF THE MORPHOLOGICAL FIDELITY OF GASTROPOD STEINKERNS

Abstract Gastropods are commonly preserved as steinkerns (internal casts), a mode of fossilization that leads to loss of external morphological features. This loss of information is problematic for taxonomic identification and ecological inference in evaluating assemblages where original shell material is not preserved. We seek to quantify how closely gastropod steinkerns represent the morphology of their original shells. We investigated this relationship experimentally by fabricating steinkerns in silicone from modern gastropod shells and comparing their geometry to that of the shells we used to create them. In addition to recording traces of ornamentation such as ribs and spines, we used a theoretical morphospace framework to evaluate the fidelity of shell-coiling parameters in steinkerns. Our results show that some morphotypes reflect their taxonomic identification more accurately than others, indicating that steinkern fidelity is highly variable. Experimental steinkerns consistently cluster less reliably by morphotype than their original shell counterparts. Additionally, we find that shell thickness is an important factor in determining steinkern fidelity. The fidelity of the high-spired Duplicaria duplicata, for example, is significantly lower than the average value for the morphotypes investigated whereas the fidelity of planispiral Haplotrema concavum and open-coiling Epitonium is significantly higher, a trend related to shell thickness. Thus, taxonomic identification and subsequent analyses, such as community composition, of steinkern assemblages must recognize this differential fidelity to counter preservational biases.

A staging table of Balkan crested newt embryonic development to serve as a baseline in evolutionary developmental studies.

There is an increased interest in the evolution and development of newts from the genus Triturus because: (1) morphological differentiation among the nine constituent species largely corresponds to different ecological preferences, (2) hybridization between different species pairs has various evolutionary outcomes in terms of life history traits and morphology, and (3) the genus expresses a balanced lethal system that causes arrested growth and death of half of the embryos. These features provide natural experimental settings for molecular, morphological, and life-history studies. Therefore, we produce a staging table for the Balkan crested newt (T. ivanbureschi). We provide detailed descriptions of 34 embryonic stages based on easily observable and interpretable external morphological characters, to ensure reproducibility. Compared with previous staging tables for Triturus, we include a vastly increased sample size and provide high-resolution photographs in lateral, ventral, and dorsal view, complemented by videos of specific developmental periods, and accompanied by detailed explanations on how to delineate the specific stages. Our staging table will serve as a baseline in comparative studies on Triturus newts: an emerging model system in evolutionary and developmental studies.

Integrative studies of Microtus hartingi and Microtus guentheri (Cricetidae, Arvicolinae) and taxonomic problem of Microtus elbeyli

In this study, Taxonomic differences between Microtus guentheri and Microtus hartingi, which are similar in terms of external morphological features, were determined using linear morphometric, geometric morphometric, karyological and molecular analyses methods. Through classical morphometric (mean values) and geometric morphometric analyses of craniodental and mandible characters, influenced by various genetic, developmental, and functional processes, it was found that M. hartingi and M. guentheri exhibited notable differences in both mean linear measurements and shape. The Jackknife in classical morphometric analyses of samples, which underwent molecular and karyological species diagnoses, was higher than 88 % for each character. In geometric morphometric analyses, the Procrustes distance for shape follows the order Ventral < Dorsal < Mandible < M3<M2<M1. Based on cross-validation, the accurate classification rate is between 73 and 100 %. In both mean linear measurements and centroid size, M. hartingi specimens exhibited significantly larger mandibles and craniodentals compared to individuals of M. guentheri. According to PLS results, there was a significant relationship between the shape of M1, M2 and M3 and environmental variables, while the relationship between their size and environmental variables was not significant. However, neither the shape nor the size of the other parts had an sufficient relationship with environmental variables. Both species exhibited karyological similarity with a chromosome number of 2n = 54, but display polymorphism in chromosome arm number. There were differences in the C-banding of autosomal chromosomes and X chromosome. The classification of species in classical and geometric morphometric analyses, diverges from the outcomes of karyological and molecular assessments. In both species, the intraspecific genetic distance remained below 1.35 %. The cyt-b exhibits significant differentiation between the two species (K2P = 6.55 %). The divergence time for M. guentheri from the clade containing M. hartingi + M. qazvinensis + Microtus dogramacii was estimated to be 0.73 million years ago. Since no sample of Microtus elbeyli with karyological and cyt-b characteristics was found in the samples taken from the type locality, the taxonomic status of M. elbeyli remains unclear.

Impact of Apis cerana cerana (Hymenoptera: Apidae) body size on newly built cell size

The influence of cell structure size on the morphological development of honey bees has been widely studied. However, the effect of the honey bees' body size on the cell size they build remains unclear. This study investigated the influence of the body size of Apis cerana cerana on the structure size of newly built honey cells, worker cells, and drone cells. Accordingly, we established colonies of A. c. cerana with large and small body-size workers. We then analyzed the birth weight and seven external morphological characteristics of workers, as well as the depth and diameter of their newly built honey cells, worker cells, and drone cells. We found significant differences in the birth weight, proboscis length, thorax width, right forewing length, femur length, tibia length, and metatarsus length of the right hindleg, and total length of the 3rd and 4th tergite of workers from large-body-size colonies (LBSC) and small-body-size colonies (SBSC). Also, the depth and diameter of the newly built cells by LBSC were significantly larger than those by SBSC. The body size of workers positively correlated with the size of newly built cells. This study emphasizes the influence of workers' body size on the structural dimensions of newly constructed cells, providing insights into the comb-building biology of A. c. cerana.

The Problems of DNA-Barcoding the Shads of Genus Alosa (Alosidae) of the Ponto-Caspian Basin

—Numerous studies show that species identification of representatives of the genus Alosa using various genetic markers is often difficult and the search for more specific biomarkers is required. For the first time we analyzed polymorphism of COI gene fragment of mitochondrial DNA of two representatives of this genus (A. tanaica and A. kessleri), supplemented with new data on A. immaculata, from the waters of the Ponto-Caspian basin in comparative aspect with other representatives of the herring (Clupeoidea) genera Alosa, Clupea, Clupeonella, Sprattus, and Sardinops. The main result was the conclusion that within the genus Alosa, it is not possible to identify species using the marker used. On the one hand, specimens collected from morphologically distinct individuals and identified as different species have the same haplotypes. On the other hand, samples belonging to different species differ from each other by an insignificant number of nucleotide substitutions and do not form independent clades on the phylogram and haplotype network. This indicates the absence of genetic differentiation between the studied samples of herrings of genus Alosa into separate species and species groups when using DNA barcoding based on the COI gene. The reasons for such a phenomenon may be the following: (1) incorrect identification of species in catches, since shads (Alosidae) have high morphological flexibility and in many species, the main external morphological characters often overlap; (2) recent time of speciation by the standards of biological evolution for shads of genus Alosa; (3) difference in proportion of interspecific hybrids, which can vary significantly between populations of the same species.