Tiger beetle fauna (Coleoptera: Carabidae, Cicindelinae) of Morocco: distribution, phenology and list of taxa

The diversity and distribution of Cicindelinae in Morocco, including new unpublished data, is summarized and discussed. In total 17 species are reported from the country. Cicindela campestris campestris is excluded from the Moroccan fauna while the occurrence of Myriochila mirei is doubtful and should be confirmed by new data. The area adjacent to the Mediterranean Sea and Atlantic Ocean coastlines holds the highest species diversity, while mountainous regions are characterized by lower diversity but also by high level of species endemism. Grouped on the basis of their chorotypes, Moroccan Cicindelinae fall into six different groups: West Mediterranean (44% of Cicindelinae species), Maghreb endemics (22%), Mediterranean (11%), Saharian (11%), Mediterranean-Westturanian (6%) and Afrotropico-Indo-Mediterranean (6%). According to their phenology, the Moroccan tiger beetles can be divided into three groups: 1) spring active species, 2) spring-summer active species, and 3) summer active species

Isolation and characterization of 8 microsatellite loci for the ‘‘killer shrimp’’, an invasive Ponto-Caspian amphipod Dikerogammarus villosus (Crustacea: Amphipoda)

Dikerogammarus villosus is a freshwater amphipod of the Ponto-Caspian origin recognized as one of the 100 worst alien species in Europe, having negative impact on biodiversity and functioning of the invaded aquatic ecosystems. The species has a wide ecophysiological tolerance and during the last 20 years it has rapidly spread throughout European inland waters. In consequence, it presents a major conservation management problem. We describe eight polymorphic microsatellite loci developed for D. villosus by combining a biotin-enrichment protocol and new generation 454GS-FLX Titanium pyrosequencing technology. When genotyped in 64 individuals from two locations, the loci exhibited a mean diversity of 4.87 alleles per locus (2–13). The mean observed and expected heterozygosities were, respectively, 0.439 (0.091–0.844) and 0.468 (0.089–0.843). Gametic disequilibrium was not detected for any pair of loci. The microsatellite markers will be a valuable tool in assessing the demographic processes associated with invasion of the killer shrimp from a genetic point of view

Out of the Black Sea: Phylogeography of the Invasive Killer Shrimp Dikerogammarus villosus across Europe

The amphipod Dikerogammarus villosus has colonized most of the European main inland water bodies in less than 20 years, having deteriorating effect on the local benthic communities. Our aim was to reveal the species phylogeography in the native Black Sea area, to define the source populations for the colonization routes in continental Europe and for the newly established UK populations. We tested for the loss of genetic diversity between source and invasive populations as well as along invasion route. We tested also for isolation by distance. Thirty three native and invasive populations were genotyped for mtDNA (COI, 16S) and seven polymorphic nuclear microsatellites to assess cryptic diversity (presence of deeply divergent lineages), historical demography, level of diversity within lineage (e.g., number of alleles), and population structure. A wide range of methods was used, including minimum spanning network, molecular clock, Bayesian clustering and Mantel test. Our results identified that sea level and salinity changes during Pleistocene impacted the species phylogeography in the Black Sea native region with four differentiated populations inhabiting, respectively, the Dnieper, Dniester, Danube deltas and Durungol liman. The invasion of continental Europe is associated with two sources, i.e., the Danube and Dnieper deltas, which gave origin to two independent invasion routes (Western and Eastern) for which no loss of diversity and no isolation by distance were observed. The UK population has originated in the Western Route and, despite very recent colonization, no drastic loss of diversity was observed. The results show that the invasion of the killer shrimp is not associated with the costs of loosing genetic diversity, which may contribute to the success of this invader in the newly colonized areas. Additionally, while it has not yet occurred, it might be expected that future interbreeding between the genetically diversified populations from two independent invasion routes will potentially even enhance this success

Catch fast and kill quickly: do tiger beetles use the same strategies when hunting different types of prey?

Background Tiger beetles (Coleoptera: Cicindelidae) are fast running predatory insects preying on different small insects and other terrestrial arthropods. Prey is located by sight and captured after short and fast pursuit interspersed with pause-and-look behaviour. At least some tiger beetle species can recognise the size and location of prey using memory, which probably allows them to achieve greater hunting success. Material and Methods Two eurytopic tiger beetle species known to occur in different types of habitat were used in the study: Cicindela hybrida hybrida, a very common central European beetle found even in artificial habitats such as sandy roads or gravel pits, and Calomera littoralis nemoralis, a species widely distributed in southern European countries and occurring on sandy sea beaches, in salt marshes, as well as on sandy banks of rivers and lakes. Both species are very similar in body size. Specimens used in the study were collected in the field and later tested in the laboratory. We checked whether tiger beetles use different hunting strategies when attacking prey of different sizes and abilities to escape as well as whether the sex of the studied species makes a difference in its hunting behaviour. Results The hunting strategies of both tiger beetle species consist of the following main phases: identification, pursuit (often with stops), attack, and optional release of the prey, and then the secondary attack, abandonment of the prey, or consumption of the prey. Considerable differences were noticed in hunting behaviour depending on the type of prey, its movement ability and escape potential. Caterpillars were attacked without pursuit, in the head or directly behind the head where a concentration of nerves and main muscles responsible for walking are located. Effective attacks on beetles were executed at the connection between the thorax and the abdomen. Calomera littoralis strongly preferred slow moving prey, while Cicindela hybrida preferred in equal measure slow moving prey and medium-sized fast moving prey. The experiment on the preferred size of prey indicated small beetles and small caterpillars as favoured by Calomera littoralis, while Cicindela hybrida preferred medium-sized fast moving prey and large caterpillars. Discussion The hunting behaviour of Calomera littoralis and Cicindela hybrida is complicated and includes a number of phases allowing to locate, capture and kill the prey. Beetles are able to discriminate between different types of prey and apply different behavioural tactics to hunt it. As the particular strategies are used to increase hunting success, and as a result allow to accumulate energy for future activity of the predator, it can be expected that such a type of hunting behaviour is characteristic also of other tiger beetle species

Mitochondrial Genomes, Phylogenetic Associations, and SNP Recovery for the Key Invasive Ponto-Caspian Amphipods in Europe

The Ponto-Caspian region is the main donor of invasive amphipods to freshwater ecosystems, with at least 13 species successfully established in European inland waters. Dikerogammarus spp. and Pontogammarus robustoides are among the most successful, due to their strong invasive impact on local biota. However, genomic knowledge about these invaders is scarce, while phylogeography and population genetics have been based on short fragments of mitochondrial markers or nuclear microsatellites. In this study, we provide: (i) a reconstruction of six mitogenomes for four invasive gammarids (D. villosus, D. haemobaphes, D. bispinosus, and P. robustoides); (ii) a comparison between the structure of the newly obtained mitogenomes and those from the literature; (iii) SNP calling rates for individual D. villosus and D. haemobaphes from different invasion sites across Europe; and (iv) the first time-calibrated full mitogenome phylogeny reconstruction of several Ponto-Caspian taxa. We found that, in comparison to other gammarids, the mitogenomes of Ponto-Caspian species show a translocation between the tRNA-E and tRNA-R positions. Phylogenetic reconstruction using the mitogenomes identified that Ponto-Caspian gammarids form a well-supported group that originated in the Miocene. Our study supports paraphyly in the family Gammaridae. These provided mitogenomes will serve as vital genetic resources for the development of new markers for PCR-based identification methods and demographic studies

Morphology and genome size of Epipactis helleborine (L.) Crantz (Orchidaceae) growing in anthropogenic and natural habitats

Background The process of apophytism or spreading native species to human-made habitats is one of the main elements in the creation of plant cover in anthropogenic areas. Lately, an increase of anthropogenic localities with valuable flora has been observed. Apophytes are also members of the family Orchidaceae, especially from the genus Epipactis. The aim of the study was to (i) determine and compare the phenotypic variation of E. helleborine (L.) Crantz plants in anthropogenic and natural habitats, (ii) compare the genome size of plants growing in natural and anthropogenic habitats. The results reported in this study may indicate that a habitat influences morphological characteristics of plant species. Methods Field studies were conducted on four native stands and four stands in anthropogenic areas of E. helleborine in Poland in years 2011–2013. Biometrical analyses were performed on shoots and flowers. The flowers were characterised by 25 biometric features and measured using a Nikon SMZ 800 binocular, microscopic Moticam-1SP cameras and the MIPlus07 programme (Conbest Co.). The nuclear DNA content was determined in fresh and young leaves of E. helleborine, collected from four natural and four anthropogenic populations. Results We observed that in anthropogenic populations: (i) shoots were higher than shoots from natural populations, (ii) flowers differed significantly in terms of ten biometric features between habitats, (iii) the genome size of some population differed significantly between plants growing in natural and anthropogenic habitats. Discussion According to some researchers, the presence of phenotypic variability and the occurrence of ecotypes are adaptation strategies of plants to environmental changes. In our opinion, in the case of the studied anthropogenic habitats (roadside) in which the E. helleborine populations grew, we can talk about ecofen due to the often repeated set of characteristic features, i.e., high shoots, long inflorescence and long, broad leaves. We agree, however, that it is difficult to isolate a taxonomic unit for ecofen due to the lack of experimental research

Cenozoic origins of the genus Calliarcys (Insecta, Ephemeroptera) revealed by Micro‑CT, with DNA barcode gap analysis of Leptophlebiinae and Habrophlebiinae

We are grateful to Michel Sartori (Lausanne, Switzerland) and José Ángel Martín del Arco (Salamanca, Spain) for donating the specimens of C. humilis to SMNS, BC CAS, and UŁ collections. We are also grateful to Christel and Hans-Werner Hoffeins (Hamburg, Germany), Mike Reich (BSPG, Munich, Germany), and Evgeny Perkovsky (Schmalhausen Institute of Zoology, Kyiv, Ukraine) for access to their collections of fossil mayflies from the Eocene Rovno amber. We would like to thank Kateřina Bláhová (IE, BC CAS) and Milan Pallmann (SMNS) for technical assistance with line drawings and the preparation of a set of macro photographs. Tomasz Mamos (UniLodz, Poland) is acknowledged for his help in the Bayesian reconstruction of phylogeny, and Łukasz Trębicki (UniLodz, Poland) for help in molecular laboratory. Comments from reviewers helped to improve the manuscript. JA-T thank the staff of Bruker SkyScan in Kontich (Belgium) for their effectiveness and fast support, for their constant improvements to the software, and for implementing the new options we requested. In this respect, we are especially indebted to Alexander Sasov (now at NeoScan, https:// neosc an. com), Stephan Boons, Xuan Liu, Phil Salmon, and Vladimir Kharitonov. We would like to thank the reviewers for their thoughtful comments and efforts towards improving our manuscript. LSID urn:lsid:zoobank. org:pub:C58BEE82-0EC6-4C59-A02D-1E5F796179B6Additional information Supplementary Information The online version contains supplementary material available at https:// doi. org/ 10. 1038/ s41598- 022- 18234-4.Funding Open access funding enabled and organized by the University of Łódź (Poland). RJG acknowledges the financial support of the Grant Agency of the Czech Republic (No. 21-05216S) and institutional support of the Institute of Entomology (Biology Centre of the Czech Academy of Sciences) RVO: 60077344. Acquisition of research equipment used in this study has been carried out within equipment subsidy granted by Alexander von Humboldt Foundation [Georg Forster Research Fellowship for Experienced Researchers] for RJG.Mayflies (Ephemeroptera) are among the oldest pterygote insects, with the earliest fossils dating back to the Late Carboniferous. Within mayflies, Leptophlebiidae are a highly diverse and widespread group, with approximately 140 genera and 640 species. Whereas taxonomy, systematics, and phylogeny of extant Leptophlebiidae are in the focus of extensive studies, little is known about leptophlebiid fossil taxa. Because fossil remains of Ephemeroptera in sedimentary rocks are relatively rare, inclusions of mayflies in amber are a unique source of information on their evolution and diversity in the past. Leptophlebiidae found in Cenozoic resins mostly belong to the subfamilies Leptophlebiinae (in Eocene Baltic amber) and Atalophlebiinae (in Miocene Dominican and Mexican ambers). In the present contribution, we confirm the first finding of the genus Calliarcys from Eocene Baltic amber by using Micro-CT, which allowed confirming its generic placement by visualizing diagnostic key characters otherwise hidden by a cloud of turbidity. Additionally, we present first molecular data on the extant species Calliarcys humilis Eaton, 1881 from the Iberian Peninsula and the barcode gap analysis for Leptophlebiinae and Habrophlebiinae.Funding Open access funding enabled and organized by the University of Łódź (Poland)Grant Agency of the Czech Republic (No. 21-05216S)Institute of Entomology (Biology Centre of the Czech Academy of Sciences) RVO: 60077344Alexander von Humboldt Foundation [Georg Forster Research Fellowship for Experienced Researchers

Morphometric traits in the fine-leaved fescues depend on ploidy level: the case of Festuca amethystina L.

Background Polyploid specimens are usually characterized by greater exuberance: they reach larger sizes and/or have a larger number of some organs. Festuca amethystina L. belongs to the section Aulaxyper. Based on morphological features, four subspecies of F. amethystina have been already identified. On the other hand, it has two cytotypes: diploid and tetraploid. The main aim of our study was to distinguish morphological differences between the cytotypes of F. amethystina, assuming that its phenotype differs significantly. Methods The nuclear DNA content was measured by flow cytometry in dry leaves from specimens originating from 13 populations of F. amethystina. Several macrometric and micrometric traits of stems, spikelets and leaf blades were taken into account in the comparative analysis of two cytotypes. Results In the case of cytotypes, specimens of tetraploids were larger than diploids. The conducted morphometric analysis of leaf cross-sections showed significant differences between the cytotypes. Discussion The research has confirmed for the first time that in the case of F. amethystina the principle of greater exuberance of polyploids is true. Differences between the cytotypes are statistically significant, however, they are not enough to make easy the distinction of cytotypes on the basis of the measurements themselves. Our findings favor the rule known in Festuca taxonomy as a whole, i.e. that the ploidy level can be one of the main classification criteria

Filogeografia inwazyjnego obunoga Dikerogammarus villosus w Europie

The killer shrimp, Dikerogammarus villosus (Crustacea Amphipoda), has been recognised as one of the 100 worst alien species in Europe, representing a major issue for local biodiversity and ecosystem functioning. During the last 20 years, it has ccolonized Central and Western Europe via the central and southern invasion corridors. Its spread was associated mainly with commercial shipping in large waterways but it was also transported overland to several Alpine lakes and even overseas to the British Isles. This thesis is an attempt to track the routes and dynamics of invasion of this one of the most effective freshwater invader. Molecular methods were used to reveal phylogeography of D. villosus in the native and invaded area as well as to tackle conservation issues. Through molecular studies we aimed to understand the dynamics of the species invasion in Europe. Our material consisted of about 606 individuals from its entire European range, sequenced for 16S and COI mitochondrial markers and 1228 individuals sequenced for seven microsatellite loci each. In result, four major populations in Danube, Dnieper, Dniester mouth and also Durngol Liman in Turkey were identified in the species native area. Two of them, from Dnieper and Danube mouth, were donors for independent routes of invasion along central and southern corridor. Microsatellite data allow to indicate Western Europe populations as a origin of latest introduction of Dikerogammarus villosus to UK. Strong bottleneck effects may be seen in the populations from the isolated Alpine Lakes, showing that they come from a very low number of introduced individuals (showing high risk of overland transport). Such complex knowledge on the phylogeography of the killer shrimp may help to decrease risk of its future introductions to other areas, such as the North American Great Lakes and Balkan lakes.Inwazyjny obunóg Dikerogammarus villosus (Crustacea Amphipoda), został uznany za jednego z 100 najbardziej ekspansywnych obcych gatunków w Europie i stanowi poważne zagrożenie dla lokalnej różnorodności biologicznej i funkcjonowania ekosystemów. W ciągu ostatnich 20 lat, gatunek ten skolonizował znaczny obszar Europy Środkowej i Zachodniej poprzez centralny i południowych korytarz inwazji Ponto-Kaspijskich hydrobiontów. Jego rozprzestrzenianie się jest związane z transportem śródlądowym dużymi drogami wodnymi, ale również został przypadkowo wsiedlony do izolowanych jezior alpejskich, a nawet na Wyspy Brytyjskie. W niniejszej pracy podejmuję próbę prześledzenia trasy i dynamiki inwazji, oraz zidentyfikowania populacji źródłowych, dla tego jednego z najbardziej skutecznych słodkowodnych gatunków inwazyjnych. Metody molekularne zostały wykorzystane do prześledzenia filogeografii D. villosus w rodzimym i nowo skolonizowanym obszarze Europy. Przeanalizowany materiał składał się z 606 osobników z 45 populacji z całej Europy, zsekwencjonowanych dla markerów mitochondrialnych 16S i i COI, oraz 1228 osobników zsekwencjonowanych dla każdego z siedmiu loci mikrosatelitarnych. Udało się ustalić cztery odmiennie genetycznie populacje w rejonie rodzimym w deltach Dunaju, Dniepru, Dniestru, a także w jeziorze Durngol w Turcji. Dwie z nich, w Dunaju i Dnieprze, były źródłami dla niezależnych dróg inwazji wzdłuż centralnego i południowego korytarza. Dane mikrosatelitarne pozwalają wskazać południowy korytarz inwazji w Europie Zachodniej jako źródło wprowadzenia Dikerogammarus villosus do Wielkiej Brytanii. Silne efekty założyciela i zubożenie puli genetycznej są wyraźnie widoczne w populacjach z izolowanym jezior alpejskich, co wskazuje na bardzo nieliczne populacje założycielskie.This study was supported by the Polish Ministry of Science and Higher Education and National Science Centre (projects no. N N304 350139 and 2011/03/D/NZ8/03012) and partially by the internal funds of the University of Lodz. Part of the work was done during scientific internship in Equipe Ecologie Evolutive, Biogeosciences, Universite de Bourgogne, France founded by the ERASMUS student exchange program and aforementioned projects