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We studied the highly abundant chromoviral Tekay clade in species from three sister genera - Anemone, Pulsatilla and Hepatica (Ranunculaceae). With this clade, we performed a concomitant survey of its phylogenetic diversity, chromosomal organisation and transcriptional activity in Anemone s.l. in order to investigate dynamics of the Tekay elements at a finer scale than previously achieved in this or any other flowering clade. The phylogenetic tree built from Tekay sequences conformed to expected evolutionary relationships of the species; exceptions being A. nemorosa and A. sylvestris, which appeared more closely related that expected, and we invoke hybridisation events to explain the observed topology. The separation of elements into six clusters could be explained by episodic bursts of activity since divergence from a common ancestor at different points in their respective evolutionary histories. In Anemone s.l. the Tekay elements do not have a preferential position on chromosomes, i.e. they can have a: (i) centromeric/pericentromeric position; (ii) interstitial position in DAPI-positive AT-rich heterochromatic regions; can be (iii) dispersed throughout chromosomes; or even (iv) be absent from large heterochromatic blocks. Widespread transcriptional activity of the Tekay elements in Anemone s.l. taxa indicate that some copies of Tekay elements could still be active in this plant group, contributing to genome evolution and speciation within Anemone s.l. Identification of Tekay elements in Anemone s.l. provides valuable information for understanding how different localisation patterns might help to facilitate plant genome organisation in a structural and functional manner. [ABSTRACT FROM AUTHOR]

This work examines the bioaccumulated organochlorine compounds in Muraena helena to draw attention to this species as a potential bioindicator organism. The morays were caught in the East Adriatic Sea, Elaphite Islands, near Dubrovnik, Croatia (lat: 42°45′38.8″; long: 17°45′53.6″) at the same location in summer (August, 2009) and winter (January, 2010), at depths from 5 to 10 m. Nine fish were caught by longline hooks per each field trip (total in summer and winter, N = 18). No single fish was free of organochlorines ( OC), but not all pesticides were present in every fish. The concentration level was: Endrin > pp DDE > Heptachlor epoxide > Heptachlor > Aldrine > Lindane > β HCH > δ HCH > Endrin aldehyde). Endrin was the only pesticide approaching the maximum concentrations proposed by the European Union and Croatia (summer max. concentration = 14.75 μg kg−1; winter max. concentration = 8.35 μg kg−1). All other pesticides ranged from 0.1 to 0.7 μg kg−1. Heptachlor was absent in winter and δ- HCH and Endrin aldehyde in summer, while Lindane and pp- DDE had higher (P ≤ 0.05) concentrations in summer. Other residues had similar seasonal concentration levels. The p- DDT was similar as in other biota from the Adriatic or Mediterranean. HCH concentrations corresponded to those found in organisms from the Italian area of the Adriatic. The endocrine and reproductive disrupting potential of the detected OCs are bases for future studies. Results show that morays are a good bioindicator model, with slight interspecies and seasonal variability in OC bioaccumulation as a consequence of diverse food consumption. [ABSTRACT FROM AUTHOR]

Only two leaf-like pygmy grasshopper species and specimens are known from Madagascar: the Leatherback Pygmy Grasshopper ( Lepocranusfuscus Devriese, 1991) -which has a relatively low median carina of the pronotum; and the Malagasy Litterhopper ( Valalyllumfolium gen. et. sp. nov. ), herein described - which has a high median carina. Lepocranusfuscus is known from the rainforests around Tampolo, Manakambahiny, and Mahavelona (Foulpointe). The new taxon, Valalyllumfolium , is described for the two mentioned genera because its members are different from the Caribbean leaf-like Choriphyllini Cadena-Castañeda & Silva, 2019, from the African leaf-like Xerophyllini Günther, 1979, and from the Asian leaf-like Cladonotini Bolívar, 1887. A tabular key to the tribes of Cladonotinae with leaf-like representatives is provided, together with photographs of type specimens of both species belonging to the newly described tribe. The holotype of the new species belongs to the Muséum national d'Histoire naturelle Orthoptera collection, Paris.gen. et. sp. nov. is known only from the Belanono forest. Both species inhabit northeastern Madagascar. The new species could be rare or not-easy-to-spot in the rainforest leaf litter, where it most probably lives. A new tribe, Valalyllini trib. nov. , is described for the two mentioned genera because its members are different from the Caribbean leaf-like Choriphyllini Cadena-Castañeda & Silva, 2019, from the African leaf-like Xerophyllini Günther, 1979, and from the Asian leaf-like Cladonotini Bolívar, 1887. A tabular key to the tribes of Cladonotinae with leaf-like representatives is provided, together with photographs of type specimens of both species belonging to the newly described tribe. The holotype of the new species belongs to the Muséum national d'Histoire naturelle Orthoptera collection, Paris., (Maks Deranja, Niko Kasalo, Karmela Adžić, Damjan Franjević, Josip Skejo.)

Modern accounts of eukaryogenesis entail an endosymbiotic encounter between an archaeal host and a proteobacterial endosymbiont, with subsequent evolution giving rise to a unicell possessing a single nucleus and mitochondria. The mononucleate state of the last eukaryotic common ancestor (LECA) is seldom, if ever, questioned, even though cells harboring multiple (syncytia, coenocytes, and polykaryons) are surprisingly common across eukaryotic supergroups. Here, we present a survey of multinucleated forms. Ancestral character state reconstruction for representatives of 106 eukaryotic taxa using 16 different possible roots and supergroup sister relationships, indicate that LECA, in addition to being mitochondriate, sexual, and meiotic, was multinucleate. LECA exhibited closed mitosis, which is the rule for modern syncytial forms, shedding light on the mechanics of its chromosome segregation. A simple mathematical model shows that within LECA's multinucleate cytosol, relationships among mitochondria and nuclei were neither one-to-one, nor one-to-many, but many-to-many, placing mitonuclear interactions and cytonuclear compatibility at the evolutionary base of eukaryotic cell origin. Within a syncytium, individual nuclei and individual mitochondria function as the initial lower-level evolutionary units of selection, as opposed to individual cells, during eukaryogenesis. Nuclei within a syncytium rescue each other's lethal mutations, thereby postponing selection for viable nuclei and cytonuclear compatibility to the generation of spores, buffering transitional bottlenecks at eukaryogenesis. The prokaryote-to-eukaryote transition is traditionally thought to have left no intermediates, yet if eukaryogenesis proceeded via a syncytial common ancestor, intermediate forms have persisted to the present throughout the eukaryotic tree as syncytia but have so far gone unrecognized., (© The Author(s) 2021. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.)

Wide nosed pygmy grasshoppers (Tetrigidae: Cladonotinae) are grasshoppers in which the frontal costa bifurcates into strongly divergent facial carinae, between which there is wide scutellum. Altogether seven genera and thirteen species inhabit India and Sri Lanka. Tribe Cladonotini is represented in this region by five genera and nine species (Cladonotus-4 spp., Deltonotus-2 spp., Gignotettix-1 sp., Hancockella-1 sp., and Yunnantettix-1 sp.), while the tribe Xerophyllini by two genera, Tettilobus (3 sp.) and Potua (1 sp.). For this Potua species it is not clear if it belongs to this genus. A simple identification key to Cladonotinae of India and Sri Lanka is provided. New synonyms are: Deltonotus humilis syn. nov., Epitettix tamilus syn. nov. and Potua aptera syn. nov., synonymous with Deltonotus gibbiceps. Deltonotus cristatus should be regarded nomen nudum. A new species, assigned to the genus Tettilobus is described from the Western Ghats of Kerala, Tettilobus trishula Skejo, Bhaskar et Stermšek sp. n. Furthermore, we present the first records of D. subcucullatus from India (Kerala).

Madagascar is home to some of the largest and most colorful pygmy grasshoppers (Tetrigidae) in the world, known as 'Malagasy Metrodorinae'. Among them, Devil's pygmy grasshoppers (genus Holocerus Bolívar, 1887) are unique in having two long spines on the back, which are modified internal lateral pronotal carinae. The genus Holocerus was composed of two species - H. lucifer (Serville, 1838) and H. taurus Rehn, 1929 syn. nov. , but here it is evidenced that the latter represents a junior synonym of the former. Simultaneously, H. devriesei sp. nov. is described as a species new to science. Holocerus lucifer is a northern species of paler coloration and longer spines (distributed from Marojejy and Maroantsetra in the north to Zahamena in the south), whereas H. devriesei sp. nov . represents the southern and darker species (distributed from Vohimana and Andasibe-Mantadia in the south to the Antongil Bay in the north). There are potential overlaps in the distribution of the two species, but without more georeferenced localities, it is impossible to discriminate whether they occur only sympatrically or also syntopically., (Josip Skejo, Kristian Medak, Marko Pavlović, Davorka Kitonić, Rafanomezanjanahary Jean Christian Miko, Damjan Franjević.)

Knowledge on the pygmy grasshoppers of Australia is, despite the numerous endemics being described from this unique continent, still scarce. Of interest is the Vingselina genus group, including genera Anaselina Storozhenko, 2019, Paraselina Storozhenko, 2019, Selivinga Storozhenko, 2019 and Vingselina Sjöstedt, 1921. The systematic position of this group, currently assigned to Batrachideinae (Bufonidini), is probably not correct. In this study new records are presented of Anaselina minor (Sjöstedt, 1921), Paraselina brunneri (Bolívar, 1887), P. trituberculata (Sjöstedt, 1932), and Selivinga tribulata Storozhenko, 2019, all except A. minor the first records of the species since their original descriptions. The first photographs of living specimens of A. minor , P. brunneri , P. trituberculata and S. tribulata are provided and their habitats described. All the records were compiled by citizen scientists who use online social media, such as iNaturalist. Lastly, P. multifora (Rehn, 1952) syn. nov . represents a junior synonym of P. brunneri ., (Josip Skejo, Matthew Connors, Michael Hendriksen, Nick Lambert, Griffin Chong, Ian McMaster, Nick Monaghan, David Rentz, Reiner Richter, Kathy Rose, Damjan Franjević.)

There are numerous pygmy grasshoppers (Tetrigidae) that exhibit leaf-like appearance. Leaf-mimic species can be found mainly in the subfamily Cladonotinae (tribes Cladonotini, Xerophyllini). Two leaf-mimic pygmy grasshopper species found in India, Pakistan (Oxyphyllum pennatum Hancock, 1909) and Borneo (Paraphyllum antennatum Hancock, 1913) are the only macropterous leaf-like species of Asia and were traditionally assigned to subfamily Cladonotinae. Our study present new records of these species, as well as updated descriptions. Oxyphyllum pennatum is recorded for the first time from Chhattisgarh (Central India), and furthermore, records from Pakistan were reviewed and confirmed. Paraphyllum antennatum is reported from a few localities in the mountains of Borneo (East Malaysia). New records found in online social media (Flickr, iNaturalist) were implemented. Brachypronotal specimens of P. antennatum are reported. After comparison with Cladonotini and Xerophyllini members, we conclude that Oxyphyllum and Paraphyllum should not be regarded Cladonotinae members. Traditional assignment was based on superficial resemblance to leaf-like Cladonotinae genera (e.g. Phyllotettix, Hymenotes, Holoarcus, Xerophyllum). However, comparison of morphological characters reveals that Oxyphyllum shares morphology with Tetriginae genera, while Paraphyllum shares morphology with Asian Metrodorinae. Here, we assign the genus Oxyphyllum to Tetriginae and the genus Paraphyllum to Metrodorinae.

The gypsy moth, Lymantria dispar , a prominent polyphagous species native to Eurasia, causes severe impacts in deciduous forests during irregular periodical outbreaks. This study aimed to describe the genetic structure and diversity among European gypsy moth populations. Analysis of about 500 individuals using a partial region of the mitochondrial COI gene, L. dispar was characterized by low genetic diversity, limited population structure, and strong evidence that all extant haplogroups arose via a single Holocene population expansion event. Overall 60 haplotypes connected to a single parsimony network were detected and genetic diversity was highest for the coastal populations Croatia, Italy, and France, while lowest in continental populations. Phylogenetic reconstruction resulted in three groups that were geographically located in Central Europe, Dinaric Alps, and the Balkan Peninsula. In addition to recent events, the genetic structure reflects strong gene flow and the ability of gypsy moth to feed on about 400 deciduous and conifer species. Distinct genetic groups were detected in populations from Georgia. This remote population exhibited haplotypes intermediate to the European L. dispar dispar , Asian L. dispar asiatica , and L. dispar japonica clusters, highlighting this area as a possible hybridization zone of this species for future studies applying genomic approaches.

The concept of species is one of the core concepts in biology and one of the cornerstones of evolutionary biology, yet it is rife with conceptual problems. Philosophers of biology have been discussing the concept of species for decades, and in doing so they sometimes appeal to the views of biologists. However, their statements as to what biologists think are seldom supported by empirical data. In order to investigate what biologists actually think about the key issues related to the problem of species, we have conducted a survey on the sample of 193 biologists from the population of biologists from over 150 biology departments at universities in the US and the EU. This article presents and discusses the results of the survey. Some results confirm and others falsify the reiterated statements of philosophers of biology as to what biologists think, but all results we obtained should be informative and relevant for future discussions of the problem of species.

In this study, the evolutionary history of the white-clawed crayfish (WCC) was evaluated using large-scale datasets comprising >1350 specimens from the entire distribution range. Using species delimitation methods on mitochondrial DNA (mtDNA) sequences, we propose four primary species hypotheses for WCC. Sequences for several nuclear regions were screened but none showed significant variation within WCC. This result favours a single secondary species hypothesis and indicates the existence of a mito-nuclear discordance in WCC. Therefore, mtDNA groups were considered only as genetic units that carry information about ancient divergences within WCC and not as taxonomic units. The reconstruction of ancestral ranges and divergence time estimates were used to link the current genetic structure with paleogeographic processes. These results showed that the emergence of mtDNA groups in WCC could be related to the Messinian Salinity Crisis, the climate cooling during the Pliocene and Pleistocene, and (paleo)shifting of the Adriatic Sea coastline in the Padanovenezian Plain. The most recent common ancestor of the mtDNA groups most likely originated from Dalmatia (eastern Adriatic coast) as indicated by the reconstruction of ancestral ranges. This ecoregion, along with the Gulf of Venice Drainages, harbours a high genetic diversity and should be emphasised as an area of the highest conservation priority., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Symbiotic associations are of broad significance in evolution and biodiversity. Green Hydra is a classic example of endosymbiosis. In its gastrodermal myoepithelial cells it harbors endosymbiotic unicellular green algae, most commonly from the genus Chlorella. We reconstructed the phylogeny of cultured algal endosymbionts isolated and maintained in laboratory conditions for years from green Hydra strains collected from four different geographical sites within Croatia, one from Germany and one from Israel. Nuclear (18S rDNA, ITS region) and chloroplast markers (16S, rbcL) for maximum likelihood phylogenetic analyses were used. We focused on investigating the positions of these algal endosymbiotic strains within the chlorophyte lineage. Molecular analyses established that different genera and species of unicellular green algae are present as endosymbionts in green Hydra, showing that endosymbiotic algae growing within green Hydra sampled from four Croatian localities are not monophyletic. Our results indicate that the intracellular algal endosymbionts of green Hydra have become established several times independently in evolution., (Copyright © 2015 Elsevier Inc. All rights reserved.)

The neon flying squid Ommastrephes bartramii Lesueur, 1821 belongs to the Ommastrephidae, Cephalopoda family. The family Ommastrephidae (Mollusca: Cephalopoda) includes many commercially important species, dispersed around the world. The Ommastrephidae family is conventionally divided into three subfamilies (Illicinae, Todarodinae, and Ommastrephinae). We report a specimen of neon flying squid caught in the winter 2013 at Luka Šipanska, Island of Šipan, Croatia and identified at the genetic level using the standard mitochondrial COI barcode region. This record represents the first genetic identification of a neon flying squid from the Adriatic Sea.

Bryozoans are aquatic invertebrates that inhabit all types of aquatic ecosystems. They are small animals that form large colonies by asexual budding. Colonies can reach the size of several tens of centimeters, while individual units within a colony are the size of a few millimeters. Each individual within a colony works as a separate zooid and is genetically identical to each other individual within the same colony. Most freshwater species of bryozoans belong to the Phylactolaemata class, while several species that tolerate brackish water belong to the Gymnolaemata class. Tissue samples for this study were collected in the rivers of Adriatic and Danube basin and in the wetland areas in the continental part of Croatia (Europe). Freshwater and brackish taxons of bryozoans were genetically analyzed for the purpose of creating phylogenetic relationships between freshwater and brackish taxons of the Phylactolaemata and Gymnolaemata classes and determining the role of brackish species in colonizing freshwater and marine ecosystems. Phylogenetic relationships inferred on the genes for 18S rRNA, 28S rRNA, COI, and ITS2 region confirmed Phylactolaemata bryozoans as radix bryozoan group. Phylogenetic analysis proved Phylactolaemata bryozoan's close relations with taxons from Phoronida phylum as well as the separation of the Lophopodidae family from other families within the Plumatellida genus. Comparative analysis of existing knowledge about the phylogeny of bryozoans and the expansion of known evolutionary hypotheses is proposed with the model of settlement of marine and freshwater ecosystems by the bryozoans group during their evolutionary past. In this case study, brackish bryozoan taxons represent a link for this ecological phylogenetic hypothesis. Comparison of brackish bryozoan species Lophopus crystallinus and Conopeum seurati confirmed a dual colonization of freshwater ecosystems throughout evolution of this group of animals.

One of the most intriguing defence strategies which aquatic organisms developed through evolution is multixenobiotic resistance (MXR). The key mediators of MXR activity are ATP-binding cassette (ABC) transport proteins. They provide resistance of aquatic organisms by binding xenobiotics and extruding them from cells in an energy-dependent manner. Since Cyprinid fish species are common target in freshwater biomonitoring programs, we have studied the presence of two main MDR/MXR efflux transporters P-glycoprotein (Pgp, Abcb1) and MRP-like protein(s) (Abcc) in the liver of five Cyprinid species: common carp, European chub, sneep, barbel, and silver prussian carp. Their presence was evaluated on the mRNA and protein level. Various pairs of primers were designed to clone homologous fragments of MXR-related genes. At the protein level, we used Western blotting with specific monoclonal antibodies against human Pgp (Abcb1, Ab C219), MRP1 (Abcc1; Ab MRPm6) or MRP2 (Abcc2; Ab M2I-4). Transcripts of both key types of MXR transporters were identified in all species examined and here we provide the phylogenetic analysis of new partial sequences. Immunochemical determinations with mammalian antibodies failed to identify the presence of MRP(s), but Pgp expression was found in all five Cyprinid species. These results support that MXR is a defence system mediated by both Pgp and MRP types of ABC transport proteins.

Symbiotic associations are of wide significance in evolution and biodiversity. The green hydra is a typical example of endosymbiosis. In its gastrodermal myoepithelial cells it harbors the individuals of a unicellular green algae. Endosymbiotic algae from green hydra have been successfully isolated and permanently maintained in a stable clean lab culture for the first time. We reconstructed the phylogeny of isolated endosymbiotic algae using the 18S rRNA gene to clarify its current status and to validate the traditional inclusion of these endosymbiotic algae within the Chlorella genus. Molecular analyses established that different genera and species of unicellular green algae could be present as symbionts in green hydra, depending on the natural habitat of a particular strain of green hydra.

Recent studies have revealed high local diversity and endemism in groundwaters, and showed that species with large ranges are extremely rare. One of such species is the cave shrimp Troglocaris anophthalmus from the Dinaric Karst on the western Balkan Peninsula, apparently uniform across a range of more than 500 kilometers. As such it contradicts the paradigm that subterranean organisms form localized, long-term stable populations that cannot disperse over long distances. We tested it for possible cryptic diversity and/or unexpected evolutionary processes, analysing mitochondrial (COI, 16S rRNA) and nuclear (ITS2) genes of 232 specimens from the entire range. The results of an array of phylogeographical procedures congruently suggested that the picture of a widespread, continuously distributed and homogenous T. anophthalmus was wrong. The taxon is composed of four or possibly five monophyletic, geographically defined phylogroups that meet several species delimitation criteria, two of them showing evidence of biological reproductive isolation in sympatry. COI genetic distances between phylogroups turned out to be a poor predictor, as they were much lower than the sometimes suggested crustacean threshold value of 0.16 substitutions per site. Most results confirmed the nondispersal hypothesis of subterranean fauna, but the southern Adriatic phylogroup displayed a paradoxical pattern of recent dispersal across 300 kilometers of hydrographically fragmented karst terrain. We suggest a model of migration under extreme water-level conditions, when flooded poljes could act as stepping-stones. In the north of the range (Slovenia), the results confirmed the existence of a zone of unique biogeographical conflict, where surface fauna is concordant with the current watershed, and subterranean fauna is not.

The aim of this work was to determine the effects of 20, 25 and 30 minute UV-irradiation periods lambda = 253.5 nm to two planarian species Dugesia tigrina (Gir.) and Polycelis felina (Daly.). In vivo, UV light effects have been reported to affect intracellular receptors and disrupt simple behaviour. The effects of UV-rays on mortality and behavior as well as morphological, cytological and histological changes in the two planarian species were assessed, and the course and the dynamics of regenerative processes were compared between them. Experimental populations of Dugesia tigrina and Polycelis felina species were maintained in laboratory conditions at room temperature. Mortality, behavioral and morphological changes were monitored daily by means of a light stereomicroscope. For cytological and histopathological analysis, planarians were fixed in Bouine fixative on the first, second, third, fifth and seventh day after exposure to UV-irradiation, respectively. They were embedded in paraffin, cut on a microtome, stained with toluidin blue and embedded in Canada-balsam. UV-rays caused mortality, behavioral, morphological, cytological and histological changes in each planarian species. In regeneration of damaged body parts reticular cells and neoblasts played the main role. Neoblasts as totipotent cells extremely increased in number in the area of damaged tissue, immediately after UV-exposure. Dugesia tigrina was more sensitive to UV-rays than Polycelis felina due to possession of less pigmented cells. The course of regeneration in both species was similar. Most individuals of both species regenerated in 5 to 12 days after UV-irradiation.

Highly abundant satellite DNA named TBREV is detected and characterized in the beetle Tribolium brevicornis (Insecta: Coleoptera). An outstanding peculiarity of the TBREV satellite monomer is its complex structure based on the two approximately 470-bp-long subunits, inversely oriented within a 1061-bp-long monomer sequence. The proposed evolutionary history demonstrates a clear trend toward increased complexity and length of the TBREV satellite monomer. This tendency has been observed on three levels: first as direct and inverted duplications of short sequence motifs, then by inverse duplication of the approximately 470-bp sequence segment, and, finally, by spread of inversely duplicated elements in a higher-order register and formation of extant monomers. Inversely oriented subunits share a similarity of 82% and have a high capacity to form a thermodynamically stable dyad structure that is, to our knowledge, the longest ever described in any satellite monomer. Analysis of divergences between inversely oriented subunits shows a tendency to a further reduction in similarity between them. Except in its centromeric localization, the TBREV satellite does not show similarity to other known Tribolium satellites, either in nucleotide sequence or in monomer length and complexity. However, TBREV shares common features of other Tribolium satellites that might be under functional constraints: nonconstant rate of evolution along the monomer sequence, short inverted repeats in the vicinity of an A+T tract, nonrandom distribution of A or T >/=3 tracts, and CENP-B box-like motifs. Although long inverted subunits might reinforce structural characteristics of the satellite monomer, their nucleotide sequence does not seem to be under constraints in order to preserve the dyad structure.

The results of preliminary karyological investigations of two geographically distant and morphologically slightly different Polycelis felina (Daly.) populations from central Croatia are reported. The results have shown that individuals of both populations are diploids, with the same chromosome number in their neoblasts (2n=18). Their karyotypes were composed of nine chromosome pairs, three of them are metacentric and the other six are submetacentric. Statistical evaluation of data indicated that both populations of Polycelis felina (Daly.), despite minor differences observed between them, belong to the same karyological biotype.