On the northernmost distribution of leaf beetles of the genus <Emphasis Type="Italic">Oreomela</Emphasis> (Coleoptera,Chrysomelidae): New species from mountains of southern Siberia

Three new species of leaf beetles are described: Oreomela dudkorum sp. n. (Terektinskii Ridge, Central Altai Mountains), O. tuvensis sp. n. (mountains of southeastern Republic of Tuva), and O. romantsovi sp. n. (Western Sayan Mountains). The representatives of this genus, and especially of the nominotypical subgenus, were found in Russia and in the territories mentioned for the first time. Thus, the distribution boundary of this genus is shifted 3–4° northwards. The representatives of Oreomela have most probably migrated from Tien Shan to the mountains of southern Siberia via the Mongolian Altai Mountains.     

Butterflies (Lepidoptera,Rhopalocera) of West Altai

A total of 176 species of butterflies have been found in West Altai. Their distribution ranges, phenology, biological preferences, and taxonomic structure are analyzed. Numerous Siberian butterfly species from the genera Erebia, Oeneis, Clossiana, Boeberia, and Colias have been found in West Altai which may be accounted for its westernmost position in the mountain system of southern Siberia and very humid climate. At the same time, many western Palaearctic as well as few Middle Asian butterflies have been discovered in West Altai for the first time. West Altai is the region where pairs of sister species which diverged in the Western and Eastern Palaearctic can be found in secondary contact. In this study, a zone of stable secondary sympatry of Melitaea didyma (western element) and M. latonigena (eastern element) as well as that of Maculinea arion (western element) and M. cyanecula (eastern element) has been found. These findings provide the first strong evidence for the distinctness of the species in these sister pairs.     

Variability of metric and qualitative traits of <Emphasis Type="Italic">Nitraria</Emphasis> species in relation to ecological and climatic conditions of Siberian habitats

The metric traits of shrubs, leaves, seeds, and flower parts have been studied in 19 Siberian populations of Nitraria sibirica Pall. and N. schoberi L. According to the obtained data, N. schoberi is characterized by larger shrubs (150–179 cm in height), leaves (15–30 mm in length, 5.7–8.8 mm in width), and seeds (up to 10 mm in length) and has rather wide (~2.5 mm) corolla petals. The size of anthers exceeds that of N. sibirica 1.2–1.5 times. The height of N. sibirica shrubs does not exceed 80 cm; the length of lamina, seed, and petals of corolla is 8–10, 3.7–7.4, and ~3.5 mm, respectively; and the size of anthers is 0.5–1 mm. Irrespective of habitat conditions, the stability of metric traits typical for both species has been shown to be rather high. The only dependence on climatic parameters is observed for the height of N. sibirica, which forms creeping forms in continental regions, especially in the mountains of the Altai Republic. Vegetative organs (laminas and young shoots) of N. sibirica are always more pubescent than those of N. shoberi.     

Ecological and molecular genetic features of <Emphasis Type="Italic">Ixodes persulcatus</Emphasis> and <Emphasis Type="Italic">I. pavlovskyi</Emphasis> ticks in Southern West Siberia

Ixodiae ticks have been counted and sampled in six distant locations within the Altai-Sayan Mountain Area and northern forest steppe in 2012. In total, 1829 individuals of I. persulcatus and 719 of I. pavlovskyi ticks were collected by flagging. I. persulcatus is found in all six examined locations and I. pavlovskyi is found in Western Altai, Northeastern Altai, and Northern Altai, as well as in forest parks near Akademgorodok of Novosibirsk. In total, 16S rRNA and COI sequences have been determined in 144 and 164 I. persulcatus individuals, as well as in 60 and 55 I. pavlovskyi. Selectively equivalent mutant forms are present in I. pavlovskyi populations of Northeastern Altai and Northern Altai. The hydrothermal regime in the forest parks near Novosibirsk is optimal for the I. pavlovskyi ticks. These ticks have recently colonized the forest parks near Novosibirsk and therefore are genetically close to I. pavloskyi from the Altai–Sayan Mountain Area populations.     

Preadaptation of <Emphasis Type="Italic">Formica aquilonia</Emphasis> and <Emphasis Type="Italic">Formica lugubris</Emphasis> ants (Hymenoptera,Formicidae) to low wintering temperatures in Northeast Asia

The ants Formica aquilonia and F. lugubris which inhabit the entire forest zone of the North Palaearctic and are absent from the basins of the Yana, Indigirka, and Kolyma rivers were found in the coastal area of the Sea of Okhotsk. A possible climatic conditionality of their occurrence in the Northeast is considered based on the data on the biotopic distribution of ants, the temperature causation of their overwintering, and cold hardiness. On the Sea of Okhotsk coast, these ants overwinter at a depth of 40–200 cm in the soil. During winter, the minimum soil temperature at a depth of 40 cm under the anthill was ?5°C. The supercooling temperature of F. aquilonia was not lower than ?20.2 ± 0.5°C, that of F. lugubris, not lower than ?19.6 ± 0.4°C. Half of F. aquilonia individuals did not survive the daily exposure at ?13°C, F. lugubris, at ?16°C. These two cold-resistant species could inhabit some biotopes of the Kolyma River basin, similar to F. exsecta, F. lemani, and F. sanguinea, but they are absent there for some reasons that are not related to the temperature. A similar cold hardiness is characteristic of F. aquilonia in Estonia (Maavara, 1971, 1985), where it represents a side effect of diapause, since excessive cold hardiness has no adaptive significance for insects overwintering in the non-freezing soils of Estonia. Colonization of Siberia by ant species turned out to be possible only due to the existing cold hardiness, i.e. preadaptation to low temperature. On the Sea of Okhotsk coast, cold hardiness of the ants is non-adaptive due to the relatively mild conditions during winter.     

Salicaceae-Feeding Leaf-Mining Insects in Siberia: Distribution,Trophic Specialization,and Pest Status

This paper provides an overview of the leaf-mining insect community feeding on willows (Salix spp.) and poplars (Populus spp.) in Siberia. According to published data and our own observations, 50 leaf-mining insect species (i.e., 24 species of Lepidoptera, 15 Coleoptera, 6 Diptera, and 5 Hymenoptera) feed on those two plant genera in Siberia. Using an integrative approach combining field work, morphological and DNA barcoding analyses, we identified 32 leaf-mining insect species from 14 regions across Siberia (i.e. 64% of all leaf-mining species known on Salicaceae in this part of Russia). Among them, 26 species most often found in parks and botanical gardens, represented new faunistic records for several poorly explored regions of Siberia. We have more than doubled the list of Salicaceae-feeding leaf-mining insects in Tomsk oblast, Altai krai, and the Republic of Tuva, and for the first time provided data on leaf-miners for the Khanty–Mansi Autonomous Okrug. The micromoth Phyllocnistis gracilistylella (Gracillariidae), recently described from Japan, was found on a new host plant (Salix caprea) in the south of Krasnoyarsk krai, is new for Russia. Eight leafmining insect species (i.e., five gracillariids: Phyllocnistis labyrinthella, Ph. unipunctella, Phyllonorycter apparella, Ph. sagitella, and Ph. populifoliella; two beetles: Zeugophora scutellaris and Isochnus sequensi; and one sawfly: Heterarthrus ochropoda) can outbreak on poplars, most often in urban plantations, botanical gardens, and plant nurseries in Siberia, and can also affect natural stands. Forty-five species of 50 leaf-mining insects known to feed on willow and poplar in Siberia also occur in Central and Eastern Europe. The remaining five species (Phyllocnistis gracilistylella, Phyllonorycter sibirica, Heterarthrus fasciatus, Tachyerges dauricus, and Isochnus arcticus) are recorded in Asia only. Species richness of the family Gracillariidae, the most diverse on Salicaceae in Siberia, displays 80% similarity to that in the European part of Russia and 71% to the Russian Far East. We discuss the faunal similarity of these regions and highlight the importance of applying an integrative approach combining ecological, morphological analyses, and DNA barcoding to explore and characterize the insect fauna of poorly studied regions of Asian part of Russia.     

Invasion of Siberian Pine Populations in Mountain Tundra in the Northern Urals

A new method for determining the upper forest border (UFB) as the border of the “root-closed” stand and regrowth as its potential vanguard is proposed. The mass dispersal of Pinus sibirica populations by the nutcracker (Nucifraga caryocatactes) in the UFB ecotone in the Northern Urals (Pavdinskii Kamen’ and Tretii Bugor Mountain) from the middle-mountain taiga to the mountain forest tundra and tundra at a distance reaching 1 km is found. It is done based on the original reconstruction method of the dynamics of population number in P. sibirica annual seedlings and on an analysis of their relations with the dynamics of the seed bearing and population number of the nutcracker. The trustworthy consortive relations of the number of Pinus sibirica generations in the mountain forested tundra and tundra zones, with its seed bearing and nutcracker number in the previous year in the middle-mountain taiga zone, are found. A significant increase, 3.7 times, in the number of seedling generations during the last 49 years is found in connection with the increase in the summer air temperature in the subarctic to 2.0°C. A hypothesis of the formation of the united genetic metapopulation of P. sibirica in the forest–tundra mountain ecotone zone as a consequence of its seed ornitochory from the highly different populations is formulated. A forecast of the mosaic formation of the vanguard “thin forests” in 20–25 years and the root-closed curtains of the forest in 40–50 years in the mountain tundra of the Northern Urals at the current rate of the rise in temperature is made.     

New dinocyst species (family Ceratiaceae Willey et Hickson) from the Lower Cretaceous of Southwestern Western Siberia

New species of Aptian dinoflagellate cyst of the family Ceratiaceae, Endoceratium immarinum sp. nov. and Nyktericysta (Hastodinium) sibirica sp. nov., are described. These taxa interpreted as nonmarine expand the palynological characteristics of the Vikulovo Formation of Western Siberia.     

<Emphasis Type="Italic">Talaromyces heiheensis</Emphasis> and <Emphasis Type="Italic">T. mangshanicus</Emphasis>, two new species from China

Two new species, Talaromyces heiheensis from rotten wood and T. mangshanicus isolated from soil, are illustrated and described as new to science in sections Trachyspermi and Talaromyces. The phylogenetic positions of the two new species inferred from the internal transcribed spacer, beta-tubulin, calmodulin and RNA polymerase II second largest subunit regions were carried out. Talaromyces heiheensis is phylogenetically closely related to T. albobiverticillius, T. rubrifaciens, T. solicola and T. erythromellis, and characterised by slow growth on Czapek yeast autolysate agar at 25 °C, orange conidia en masse on malt extract agar at 25 °C, biverticillate and terverticillate conidiophores, acerose phialides and subglobose to ellipsoidal, smooth-walled conidia. Talaromyces mangshanicus is related to T. kendrickii, T. qii and T. thailandensis, and characterised by slow-growing colonies with absent or sparse sporulation on CYA agar at 25 °C, conidia en masse greyish purple, purplish red soluble pigment on yeast extract agar (YES) at 25 °C, biverticillate conidiophores, ampulliform phialides and subglobose to ellipsoidal conidia with echinulate walls. They are distinguished from the known species in culture characteristics on four standard media, microscopic features and sequence data.     

Quantum Speciation of <Emphasis Type="Italic">Yersinia pestis</Emphasis> Plague Microbe in a Heteroimmune Environment: In the Populations of Hibernating Tarbagan Marmots (<Emphasis Type="Italic">Marmota sibirica</Emphasis>)

This review is devoted to the ecological mechanism for the transformation of the population of the psychrophilic saprozoobiont pseudotuberculosis microbe Yersinia pseudotuberculosis O:1b into the population of the obligate blood plague microbe Yersinia pestis in the host–parasite system of the marmot flea (Marmota sibirica–Oropsylla silantiewi). The correspondence of this mechanism to the evolutionary principle of quantum speciation has been shown. The most significant population–genetic microbial transformations took place in the populations of hibernating marmots. The main factor of rapid speciation was the heterothermic and, accordingly, heteroimmune state of host marmots during hibernation. During winter awakenings, the body temperature of marmots increases in an S-shaped manner from 5 to 37°C within a short period of time (from dozens of minutes to several hours). A drastic acceleration of metabolic and immune processes occurs in the temperature range of 20–30°C. Rapid adaptation to the “explosive” increase in the immune activity of the primary host (tarbagan marmot) during its regular winter awakenings was the essence of the process of Y. pestis speciation.     

Dynamics of the range of <Emphasis Type="Italic">Artemisia</Emphasis> genus over the territory of Western Siberia and adjacent regions in holocene in connection with climate change on the basis of pollen data

Reconstruction of the dynamics of the range of Artemisia genus was carried out with the help of palynoareological method on the basis of the data of 97 pollen diagrams from the territory of Western Siberia and the Altai-Sayan mountainous region over the millennial time sections since the Late Glacial time till the present time. It was revealed that the maximal spread of the Artemisia steppe communities (probably of the cryoarid pattern) occurred at the territory under examination at 13–10 ka yr BP. After the time point of 9 ka yr BP a fundamental change in landscapes of Western Siberia occurred which resulted in the disintegration of previously continuous Artemisia steppe cover into separate blocks. Since 8 ka yr BP the Artemisia steppe communities of modern appearance occupied the zones of modern steppe and forest-steppe of Western Siberia and Northern Kazakhstan while over the taiga zone the Artemisia plant communities were spread occasionally under the extra zonal conditions and as the pioneering plant groupings on naturally disturbed soils. Since the time point of 4 ka yr BP till the present time the northern boundary of the steppe zone under-went pulsing shifts to the north and back and also broadening and narrowing of isolated insular steppes in Altai-Sayan mountain region took place. At present time we observe an increase in the role of plants of Artemisia genus in the tundra zone of Western Siberia and in the Altai mountains.     

Nucleotide polymorphisms of candidate genes of adaptive significance in the ural populations of <Emphasis Type="Italic">Larix sibirica</Emphasis> Ledeb.

The objectives of conservation and sustainable forest management require in depth study of genomes of woody plants and definition of their intraspecific genetic diversity. In recent years, an approach was developed based on the study of “candidate genes” that can potentially be involved in the formation of adaptive traits. In this study, we investigated nucleotide polymorphism of several adaptive candidate genes in the populations of Siberian larch (Larix sibirica Ledeb.) in the Urals. Representatives of this genus are among the most valuable and widely distributed forest tree species in Russia. From ten selected gene loci in the genome of L. sibirica, we isolated and investigated three loci, one of which (ABA-WDS) was sequenced in L. sibirica for the first time. The total length of the analyzed sequence in each individual amounted to 2865 bp. The length of locus alignment was from 360 bp to 1395 bp. In total, we identified 200 polymorphic positions. The most conservative is locus 4CL1-363, and the most polymorphic is locus sSPcDFD040B03103-274. The studied populations of L. sibirica are characterized by a high level of nucleotide polymorphism in comparison with other species and genuses (Picea, Pinus, Pseudotsuga, Abies) conifers plants (Hd = 0.896; π = 0.007; θ_W = 0.015). The most selectively neutral polymorphism (D _T =–0.997) was attributed to locus 4CL1-363, and polymorphism with high probability of adaptability (D _T =–1.807) was determined for the ABA-WDS locus. We identified 54 SNP markers, only five of which were nonsynonymous (9.26%) replacements. The average frequency of SNPs in the three studied loci of L. sibirica was one SNP in 53 bp. We detected unique SNP markers for eight populations, which could potentially be used to identify populations. Populations that are characterized by the highest number of unique SNP markers can be recommended for selection in order to preserve the gene pool of the species.     

<Emphasis Type="Italic">Burkholderia alba</Emphasis> sp. nov., isolated from a soil sample on Halla mountain in Jeju island

A rod-shaped, round and white colony-forming strain AD18^T was isolated from the soil on Halla mountain in Jeju Island, Republic of Korea. Comparative analysis of 16S rRNA gene sequence revealed that this strain was closely related to Burkholderia oklahomensis C6786^T (98.8%), Burkholderia thailandensis KCTC 23190^T (98.5%). DNA-DNA relatedness (14.6%) indicated that the strain AD18^T represents a distinct species that is separate from B. oklahomensis C6786^T. The isolate grew at pH 5.0–9.0 (optimum, pH 7.0), 0–3% (w/v) NaCl (optimum, 0%), and temperature 10–40°C (optimum 35°C). The sole quinone of the strain was Q-8, and the predominant fatty acids were C_16:0, C_17:0 cyclo, and C_19:0 cyclo ω8c. The genomic DNA G + C content of AD18T was 65.6 mol%. Based on these findings, strain AD18^T is proposed to be a novel species in the genus Burkholderia, for which the name Burkholderia alba sp. nov. is proposed (= KCCM 43268^T = JCM 32403^T). The type strain is AD18^T.     

Refugium of the Boreal Forests of the Circumpolar Urals

The results of studies of the vegetative cover and ontogenetic composition of the cenopopulations of Picea obovata Ledeb. and Larix sibirica Ledeb. in the Gorno-Khadytinskii Forest Reserve, which is the refugium of the northern taiga forests of Western Siberia, have been summarized. According to the Brown–Blanke system, two associations of forests were identified and described, as well as two tundra associations, for comparison. The unique association of the Aconito septentrionalis–Piceetum obovatae taiga forests Zaugolnova et Morozova 2009 were analyzed in detail, and a comparison of these associations was carried out in the Gorno-Khadytinskii and Pechoro-Ilychskii reserves, where these forests are the reference standards of the zonal-type forests. An estimate of the ontogenetic composition of the cenopopulations of P. obovata Ledeb. and L. sibirica Ledeb. is given, as well as the absolute age of the trees of these species.     

Competition between native Antarctic vascular plants and invasive <Emphasis Type="Italic">Poa annua</Emphasis> changes with temperature and soil nitrogen availability

Over the last decades human have introduced non-native organisms to Antarctica, including the grass species Poa annua. This non-native grass under constant growth temperatures has been shown negatively affect the growth of the only two native Antarctic vascular plants, Deschampsia antarctica and Colobanthus quitensis, under constant growth temperatures. However, whether there are changes in the interaction between these species under warmer conditions is an important question. In cold ecosystems, soil nutrient status directly affects plant responses to increases in temperature and Antarctic soils are highly variable in nutrient supply. Thus, in this study we experimentally assessed the interaction between the non-native Poa with the two native Antarctic vascular plant species at two different temperatures and levels of nutrient availability. Individual mats of the study species were collected in King George Island, and then transported to Concepcion where we conducted competition experiments. In the first experiment we used soil similar to that of Antarctica and plants in competition were grown at two temperatures: 5°/2° and 11°/5 °C (day/night temperature). In a second experiment plants were grown in these two temperature regimes, but we varied nitrogen (N) availability by irrigating plants with Hoagland solutions that contained 8000 or 300 µM of N. Overall, Poa exerted a competitive effect on Deschampsia but only at the higher temperature and higher N availability. At 5°/11 °C the competitive response of Deschampsia to Poa was of similar magnitude to the competitive effect of P. Deschampsia, and the competitive effect was greater with at low N. The competitive effect of Poa was similar to the competitive response of Colobanthus to Poa at both temperatures and N levels. Thus, at low temperatures and N soil content the native Antarctic species might withstand Poa invasion, but this might change with climate warming.     

Biology and Thermal Requirements of <Emphasis Type="Italic">Fopius arisanus</Emphasis> (Sonan, 1932) (Hymenoptera: Braconidae) Reared on <Emphasis Type="Italic">Ceratitis capitata</Emphasis> Eggs (Wiedemann) (Diptera: Tephritidae)

Fopius arisanus (Sonan) is a solitary parasitoid of eggs and the first instar larvae of Tephritidae. Due to the occurrence of Ceratitis capitata (Wiedemann) in various regions and under several climatic conditions, this study aimed to evaluate the effect of different temperatures on the embryonic development (egg–adult) and determine thermal requirements and the number of annual generations F. arisanus on eggs of C. capitata. In the laboratory, eggs of C. capitata (24 h) were submitted to parasitism of F. arisanus during 6 h. Later, the eggs were placed in plastic containers (50 mL) (50 eggs/container) on a layer of artificial diet and packed in chambers at temperatures 15, 18, 20, 22, 25, 28, 30, and 32 ± 1°C, RH 70 ± 10%, and a photophase of 12 h. The largest number of offspring, emergence rate, and weight of adults of F. arisanus were observed at 25°C. The highest sex ratios (sr > 0.75) were recorded at 15 and 18°C, being statistically higher than the temperatures 20°C (0.65), 22°C (0.64), 25°C (0.65), 28°C (0.49), and 30°C (0.47). At 32°C, there was no embryonic development of F. arisanus. The egg–adult period was inversely proportional to temperature. Based on the development of the biological cycle (egg–adult), the temperature threshold (T _t) was 10.3°C and thermal constant (K) of 488.34 degree-days, being the number of generations/year directly proportional to the temperature increase. The data show the ability of F. arisanus to adapt to different thermal conditions, which is important for biological control programs of C. capitata.     

The endosymbiont <Emphasis Type="Italic">Wolbachia</Emphasis> in Eurasian populations of <Emphasis Type="Italic">Drosophila melanogaster</Emphasis>

The endosymbiotic α-proteobacteria Wolbachia is widely spread among arthropods and Filariidae nematodes. This bacterium is transmitted vertically via a transovarian route. Wolbachia is a cause of several reproductive abnormalities in the host species. We analyzed the isofemale lines created using flies collected from Drosophila melanogaster natural populations for infection with the endosymbiont Wolbachia. Wolbachia were genotyped according to five variable markers: the presence of insertion sequence IS5 in two loci, the copy number of two minisatellite repeats, and an inversion. Overall, 665 isofemale lines isolated from the populations of D. melanogaster from Ukraine, Belarus, Moldova, Caucasus, Central Asia, Ural, Udmurtia, Altai, West and East Siberia, and Far East in 1974 through 2005 were used in the work. The samples from Ukrainian, Altaian, and Middle Asian populations were largest. The infection rate of D. melanogaster populations from Middle Asia, Altaian, and Eastern Europe (Ukraine, Moldavia, and Belarus) with Wolbachia amounted to 64, 56, and 39%, respectively. The D. melanogaster population from the Caucasus displayed heterogeneity in the genotypes of this cytoplasmic infection. The Wolbachia genotype wMel, detected in all the populations studied, was the most abundant. The genotype wMelCS2 was always present in the populations from Middle Asia and Altai and was among the rare variants in the D. melanogaster populations from the Eastern Europe. Single instances of the Wolbachia genotype wMelCS occurred in a few flies from the Central Asian and Altai populations, but was not found this genotype in the other regions.     

Bacterial metabolites from intra- and inter-species influencing thermotolerance: the case of <Emphasis Type="Italic">Bacillus cereus</Emphasis> and <Emphasis Type="Italic">Geobacillus stearothermophilus</Emphasis>

Bacterial metabolites with communicative functions could provide protection against stress conditions to members of the same species. Yet, information remains limited about protection provided by metabolites in Bacillus cereus and inter-species. This study investigated the effect of extracellular compounds derived from heat shocked (HS) and non-HS cultures of B. cereus and Geobacillus stearothermophilus on the thermotolerance of non-HS vegetative and sporulating B. cereus. Cultures of B. cereus and G. stearothermophilus were subjected to HS (42 or 65 °C respectively for 30 min) or non-HS treatments. Cells and supernatants were separated, mixed in a combined array, and then exposed to 50 °C for 60 min and viable cells determined. For spores, D values (85 and 95 °C) were evaluated after 120 h. In most cases, supernatants from HS B. cereus cultures added to non-HS B. cereus cells caused their thermotolerance to increase (D ₅₀ 12.2–51.9) when compared to supernatants from non-HS cultures (D ₅₀ 7.4–21.7). While the addition of supernatants from HS and non-HS G. stearothermophilus cultures caused the thermotolerance of non-HS cells from B. cereus to decrease initially (D ₅₀ 3.7–7.1), a subsequent increase was detected in most cases (D ₅₀ 18–97.7). In most cases, supernatants from sporulating G. stearothermophilus added to sporulating cells of B. cereus caused the thermotolerance of B. cereus 4810 spores to decline, whereas that of B. cereus 14579 increased. This study clearly shows that metabolites in supernatants from either the same or different species (such as G. stearothermophilus) influence the thermotolerance of B. cereus.     

A review of sciarids of the genus <Emphasis Type="Italic">Leptosciarella</Emphasis> Tuomikoski 1960 (Diptera,Sciaridae) in the Altai fauna,with description of three new species

The taxonomy of the genus Leptosciarella Tuomikoski, 1960 in the Altai Area within Altai Territory and the Altai Republic was studied. Eleven species were recognized, including three new species which are described and illustrated here: Leptosciarella (L.) ulagana Komarova sp. n., L. (L.) angustistylus Komarova sp. n., L. (L.) senex Komarova sp. n. Additionally, L. (L.) fuscipalpa (Mohrig et Mamaev 1979) and L. (L.) truncata (Tuomikoski 1960) are recorded from Altai for the first time. A key to the Altai species is provided.