The grey wolf (Canis lupus) is an iconic large carnivore that has increasingly been recognized as... more The grey wolf (Canis lupus) is an iconic large carnivore that has increasingly been recognized as an apex predator with intrinsic value and a keystone species. However, wolves have also long represented a primary source of human¿carnivore conflict, which has led to long-term persecution of wolves, resulting in a significant decrease in their numbers, genetic diversity and gene flow between populations. For more effective protection and management of wolf populations in Europe, robust scientific evidence is crucial. This review serves as an analytical summary of the main findings from wolf population genetic studies in Europe, covering major studies from the `pre-genomic era¿ and the first insights of the `genomics era¿. We analyse, summarize and discuss findings derived from analyses of three compartments of the mammalian genome with different inheritance modes: maternal (mitochondrial DNA), paternal (Y chromosome) and biparental [autosomal microsatellites and single nucleotide polymorphisms (SNPs)]. To describe large-scale trends and patterns of genetic variation in European wolf populations, we conducted a meta-analysis based on the results of previous microsatellite studies and also included new data, covering all 19 European countries for which wolf genetic information is available: Norway, Sweden, Finland, Estonia, Latvia, Lithuania, Poland, Czech Republic, Slovakia, Germany, Belarus, Russia, Italy, Croatia, Bulgaria, Bosnia and Herzegovina, Greece, Spain and Portugal. We compared different indices of genetic diversity in wolf populations and found a significant spatial trend in heterozygosity across Europe from south-west (lowest genetic diversity) to north-east (highest). The range of spatial autocorrelation calculated on the basis of three characteristics of genetic diversity was 650-850 km, suggesting that the genetic diversity of a given wolf population can be influenced by populations up to 850 km away. As an important outcome of this synthesis, we discuss the most pressing issues threatening wolf populations in Europe, highlight important gaps in current knowledge, suggest solutions to overcome these limitations, and provide recommendations for science-based wolf conservation and management at regional and Europe-wide scales.
The ignita species group within the genus Chrysis includes over 100 cuckoo wasp species, which al... more The ignita species group within the genus Chrysis includes over 100 cuckoo wasp species, which all lead a parasitic lifestyle and exhibit very similar morphology. The lack of robust, diagnostic morphological characters has hindered phylogenetic reconstructions and contributed to frequent misidentification and inconsistent interpretations of species in this group. Therefore, molecular phylogenetic analysis is the most suitable approach for resolving the phylogeny and taxonomy of this group. We present a wellresolved phylogeny of the Chrysis ignita species group based on mitochondrial sequence data from 41 ingroup and six outgroup taxa. Although our emphasis was on European taxa, we included samples from most of the distribution range of the C. ignita species group to test for monophyly. We used a continuous mitochondrial DNA sequence consisting of 16S rRNA, tRNA Val , 12S rRNA and ND4. The location of the ND4 gene at the 3 0 end of this continuous sequence, following 12S rRNA, represents a novel mitochondrial gene arrangement for insects. Due to difficulties in aligning rRNA genes, two different Bayesian approaches were employed to reconstruct phylogeny: (1) using a reduced data matrix including only those positions that could be aligned with confidence; or (2) using the full sequence dataset while estimating alignment and phylogeny simultaneously. In addition maximum-parsimony and maximum-likelihood analyses were performed to test the robustness of the Bayesian approaches. Although all approaches yielded trees with similar topology, considerably more nodes were resolved with analyses using the full data matrix. Phylogenetic analysis supported the monophyly of the C. ignita species group and divided its species into well-supported clades. The resultant phylogeny was only partly in accordance with published subgroupings based on morphology. Our results suggest that several taxa currently treated as subspecies or names treated as synonyms may in fact constitute separate species. Our study provides a solid basis for further systematic investigations of this enigmatic insect group.
The extent of gene flow during the range expansion of non-native species influences the amount of... more The extent of gene flow during the range expansion of non-native species influences the amount of genetic diversity retained in expanding populations. Here, we analyse the population genetic structure of the raccoon dog (Nyctereutes procyonoides) in north-eastern and central Europe. This invasive species is of management concern because it is highly susceptible to fox rabies and an important secondary host of the virus. We hypothesized that the large number of introduced animals and the species' dispersal capabilities led to high population connectivity and maintenance of genetic diversity throughout the invaded range. We genotyped 332 tissue samples from seven European countries using 16 microsatellite loci. Different algorithms identified three genetic clusters corresponding to Finland, Denmark and a large 'central' population that reached from introduction areas in western Russia to northern Germany. Cluster assignments provided evidence of long-distance dispersal. Th...
Concordant differences in morphology, phenology and RAMS markers, as well as in sequenced mtDNA (... more Concordant differences in morphology, phenology and RAMS markers, as well as in sequenced mtDNA (COI, COII, cytb) and nuclear DNA (ITS2) fragments, indicate that Dolerus asper Zaddach, 1859 and Dolerus brevicornis Zaddach, 1859 are valid species. On the basis of morphology, molecular markers, and distributional records, both species are distinct from Dolerus gibbosus Hartig, 1837 (= Dolerus planatus Hartig, 1837).
... that the placement of Larentiinae may have been an artefact resulting from insufficient taxon... more ... that the placement of Larentiinae may have been an artefact resulting from insufficient taxon and character ... & Kaila (2004), as the analysis inferred Cosymbiini as the basal tribe of the ... First, feeding on Polygonaceae in the larval stage is shared by Lythriini, Rhodometrini and ...
We analyzed phylogenetic relationships of brown bear (Ursus arctos) maternal lineages in Finland ... more We analyzed phylogenetic relationships of brown bear (Ursus arctos) maternal lineages in Finland using nucleotide sequences of the mitochondrial control region of 135 individuals. A total of 14 haplotypes, which belong to 2 haplogroups, were characterized. Haplogroup I is spread across the entire bear range, whereas haplogroup D is restricted to the southeastern part of the country. Most unconventional was the concentration of haplotype C in a small territory in central Finland, which is explained by the natal philopatry of bears that descended from one or more translocated females. Analysis of Finnish brown bear haplotypes together with 15 European eastern lineage haplotypes revealed that the Romanian-Slovakian region was probably an important refuge (or route) for recolonization of Finland by brown bears after the last ice age.
... Sequences were resolved using an ABI PRISM 377 automated sequencer (Applied Biosystems, Foste... more ... Sequences were resolved using an ABI PRISM 377 automated sequencer (Applied Biosystems, Foster City, USA). ... The hypothesis of hybridization between T. griseata and T. comae receives further support from the discussion of Pittaway (1993) for closely related sphingid ...
Concordant differences in morphology, phenology and RAMS markers, as well as in sequenced mtDNA (... more Concordant differences in morphology, phenology and RAMS markers, as well as in sequenced mtDNA (COI, COII, cytb) and nuclear DNA (ITS2) fragments, indicate that Dolerus asper Zaddach, 1859 and Dolerus brevicornis Zaddach, 1859 are valid species. On the basis of morphology, molecular markers, and distributional records, both species are distinct from Dolerus gibbosus Hartig, 1837 (= Dolerus planatus Hartig, 1837). Taxonomy of the species is clarified and the neotypes of Dolerus asper Zaddach, 1859 and Dolerus brevicornis Zaddach, 1859 are designated. The synonymies of Dolerus asper Zaddach, 1859, to Dolerus planatus Hartig, 1837 and Dolerus derzavini Malaise, 1931, spec. rev. to D. asper Zaddach, 1859 are abandoned. Dolerus carbonarius Zaddach, 1859 and Dolerus fumosus Zaddach, 1859 are considered to be species inquirendae. Phylogenetic analyses of the ITS2 fragment and fragments of ITS2 + COI and ITS2 + COII yielded the topology [D. asper, (D. brevicornis, D. gibbosus)], while those of all other markers and their combinations resulted in the topology [D. brevicornis, (D. asper, D. gibbosus)]. In the latter hypothesis the clade asper + gibbosus is also supported by structural synapomorphies.
Escherichia coli ribosomes with an A to U transversion at nucleotide 1067 of their 23 S rRNA are ... more Escherichia coli ribosomes with an A to U transversion at nucleotide 1067 of their 23 S rRNA are impaired in their effective association rate constants (kcat/KM) for both EF-Tu and EF-G binding. In addition, the times that EF-G and EF-Tu spend on the ribosome during elongation are significantly increased by the A to U transversion. The U1067 mutation impairs EF-Tu function more than EF-G function. The increase in the time that EF-Tu remains bound to ribosome is caused, both by a slower rate of GTP-hydrolysis in ternary complex and by a slower EF-Tu·GDP release from the mutated ribosomes. There is, at the same time, no change in ribosomal accuracy for aminoacyl-tRNA recognition. With support from these new data we propose that nucleotide 1067 is part of the ribosomal A-site where it directly interacts with both EF-G and EF-Tu.
The raccoon dog (Nyctereutes procyonoides) is a canid that is indigenous in East Asia and alien i... more The raccoon dog (Nyctereutes procyonoides) is a canid that is indigenous in East Asia and alien in Europe, where it was introduced more than half a century ago. The aim of this study was to compare the parasite faunas associated with raccoon dogs in their native and introduced ranges, and to identify zoonotic parasite species. We examined 255 carcasses of hunted raccoon dogs from Estonia and recorded a total of 17 helminth species: 4 trematodes, 4 cestodes and 9 nematodes. The most prevalent parasite species were Uncinaria stenocephala (97.6%) and Alaria alata (68.3%). Average parasite species richness was 2.86 (the highest was 9) and only two animals were not parasitized at all. Although the infection intensity was determined by weight and not by sex, all animals infected with more than five helminth species were males. We also found that animals infected with higher numbers of helminth species fed significantly more on natural plants. Intentional consumption of grass may represent a self-medicating behaviour among raccoon dogs. We included the Estonian data into a wider comparison of raccoon dog parasite faunas and found a total of 54 helminth taxa, including 28 of zoonotic potential. In Europe, raccoon dogs are infected with a minimum of 32 helminth species of which 19 are zoonotic; in the native range they are infected with 26 species of which 17 are zoonotic. Most species were nematodes or trematodes, with fewer cestodes described. The recent increase in the number and range of raccoon dogs in Europe and the relatively high number of zoonotic parasite taxa that it harbours suggests that this species should be considered an important source of environmental contamination with zoonotic agents in Europe.
In the Baltic countries, the two zoonotic diseases, alveolar echinococcosis (AE) caused by Echino... more In the Baltic countries, the two zoonotic diseases, alveolar echinococcosis (AE) caused by Echinococcus multilocularis, and cystic echinococcosis (CE) caused by Echinococcus granulosus, are of increasing public health concern. Observations from Estonia, Latvia and Lithuania indicate that the distribution of both parasites is wider in the Baltics than previously expected. In this paper, we review and discuss the available data, regarding both parasitoses in animals and humans, from the Baltic countries and selected adjacent regions. The data are not easily comparable but reveal a worrisome situation as the number of human AE and CE cases is increasing. Despite improvements in diagnostics and treatment, AE has a high morbidity and mortality in the Baltic region. For the control of both zoonoses, monitoring transmission patterns and timely diagnosis in humans as well as the development of local control programs present major challenges.
Alveolar echinococcosis, caused by the tapeworm Echinococcus multilocularis, is one of the most s... more Alveolar echinococcosis, caused by the tapeworm Echinococcus multilocularis, is one of the most severe parasitic diseases in humans and represents one of the 17 neglected diseases prioritised by the World Health Organisation (WHO) in 2012. Considering the major medical and veterinary importance of this parasite, the phylogeny of the genus Echinococcus is of considerable importance; yet, despite numerous efforts with both mitochondrial and nuclear data, it has remained unresolved. The genus is clearly complex, and this is one of the reasons for the incomplete understanding of its taxonomy. Although taxonomic studies have recognised E. multilocularis as a separate entity from the Echinococcus granulosus complex and other members of the genus, it would be premature to draw firm conclusions about the taxonomy of the genus before the phylogeny of the whole genus is fully resolved. The recent sequencing of E. multilocularis and E. granulosus genomes opens new possibilities for performing in-depth phylogenetic analyses. In addition, whole genome data provide the possibility of inferring phylogenies based on a large number of functional genes, i.e. genes that trace the evolutionary history of adaptation in E. multilocularis and other members of the genus. Moreover, genomic data open new avenues for studying the molecular epidemiology of E. multilocularis: genotyping studies with larger panels of genetic markers allow the genetic diversity and spatial dynamics of parasites to be evaluated with greater precision. There is an urgent need for international coordination of genotyping of E. multilocularis isolates from animals and human patients. This could be fundamental for a better understanding of the transmission of alveolar echinococcosis and for designing efficient healthcare strategies.
Samples were collected and stored at 80°C for 1 week to avoid the risk of infection with any Ec... more Samples were collected and stored at 80°C for 1 week to avoid the risk of infection with any Echinococcus spp. eggs present. Approximately 250 mg was placed into 2-mL tubes, heated at 65°C for 15 min, and stored at 80°C. The heating and cooling procedure helps to break the parasite egg shells, enabling more efficient DNA extraction. DNA was extracted by using the QIAamp DNA Stool Mini Kit (QIAGEN, Hilden, Germany) according to the manufacturer's instructions. were designed to amplify short sequences of mitochondrial DNA. The Echinococcus multilocularis tapeworm-specific primer pair (EMfor1 and EMrev1) amplifies tRNA-Ile/Lys, the E. granulosus tapeworm-specific (EGfor1 and EGrev1) and fox-specific (F1 and RVu) primer pairs amplify 12S rRNA, and the dog-specific (Dog1f and HW1r) primer pair amplifies part of the mitochondrial DNA control region. Although the 3 primer pairs were highly specific, the dog primers can potentially amplify gray wolf (Canis lupus) DNA. In other similar studies, only the parasite (1) or the host species
The raccoon dog (Nyctereutes procyonoides) is an alien species in Europe and an important vector ... more The raccoon dog (Nyctereutes procyonoides) is an alien species in Europe and an important vector of zoonotic diseases. However, compared to the red fox (Vulpes vulpes), less attention has been paid to the raccoon dog as a potentially important host for Echinococcus multilocularis, the infective agent of alveolar echinococcosis, which is an emerging infectious disease with a high mortality rate. We examined the small intestines of 249 Estonian raccoon dogs and found 1.6% of individuals to be infected with E. multilocularis. The relatively large difference between this prevalence and that found in sympatric red foxes (31.5%) sampled during the same time period might be due to differences in diet: red foxes consume significantly more arvicolid rodents - the main intermediate hosts of the parasite - especially during the coldest period of the year when raccoon dogs hibernate. Nonetheless, given the relatively high density of raccoon dogs, our results suggest that the species also represents an important definitive host species for E. multilocularis in Estonia. Compared with other countries in Europe where E. multilocularis-infected raccoon dogs have been recorded (Latvia, Lithuania, Poland, Germany, and Slovakia), the prevalence in Estonia is low. The longer hibernation period of raccoon dogs at higher latitudes may explain this pattern. Both mitochondrial and nuclear loci were analysed for Estonian isolates: based on EmsB microsatellite genotyping the Estonian isolates shared an identical genotype with E. multilocularis in northern Poland, suggesting a common history with this region. The data from more than a quarter of the mitochondrial genome (3558 bp) revealed two novel haplotypes specific to Estonia and placed them into the same haplogroup with isolates from other European regions. Considering that the raccoon dog is becoming increasingly widespread and is already relatively abundant in several countries in Europe, the role of the species must be taken into account when assessing the E. multilocularis related risks to public health.
Background: Echinococcus granulosus and E. multilocularis are tapeworm parasites of major medical... more Background: Echinococcus granulosus and E. multilocularis are tapeworm parasites of major medical and veterinary importance, causing cystic and alveolar echinococcosis, respectively. Both diseases are listed among the most severe parasitic diseases in humans, representing 2 of the 17 neglected diseases prioritised by the World Health Organisation. However, little is known about the role of urban animals in transmission of both parasite species.
Previous experiments have shown that the top of helix 90 of 23S rRNA is highly important for the ... more Previous experiments have shown that the top of helix 90 of 23S rRNA is highly important for the ribosomal peptidyltransferase activity and might be part of the donor (P) site. Developing on these studies, mutations in the 23S rRNA at the highly conserved positions G2505, G2582, and G2583 were investigated. None of the mutations affected assembly, subunit association, or the capacity of tRNA binding to A and P sites. A "selective transpeptidation assay" revealed that the mutations specifically impaired peptide bond formation. Results with a modified "fragment" assay using the minimal donor substrate pA-fMet are consistent with a model where the nucleotides psiGG2582 form a binding pocket for C75 of the tRNA.
The ignita species group within the genus Chrysis includes over 100 cuckoo wasp species, which al... more The ignita species group within the genus Chrysis includes over 100 cuckoo wasp species, which all lead a parasitic lifestyle and exhibit very similar morphology. The lack of robust, diagnostic morphological characters has hindered phylogenetic reconstructions and contributed to frequent misidentification and inconsistent interpretations of species in this group. Therefore, molecular phylogenetic analysis is the most suitable approach for resolving the phylogeny and taxonomy of this group. We present a wellresolved phylogeny of the Chrysis ignita species group based on mitochondrial sequence data from 41 ingroup and six outgroup taxa. Although our emphasis was on European taxa, we included samples from most of the distribution range of the C. ignita species group to test for monophyly. We used a continuous mitochondrial DNA sequence consisting of 16S rRNA, tRNA Val , 12S rRNA and ND4. The location of the ND4 gene at the 3 0 end of this continuous sequence, following 12S rRNA, represents a novel mitochondrial gene arrangement for insects. Due to difficulties in aligning rRNA genes, two different Bayesian approaches were employed to reconstruct phylogeny: (1) using a reduced data matrix including only those positions that could be aligned with confidence; or (2) using the full sequence dataset while estimating alignment and phylogeny simultaneously. In addition maximum-parsimony and maximum-likelihood analyses were performed to test the robustness of the Bayesian approaches. Although all approaches yielded trees with similar topology, considerably more nodes were resolved with analyses using the full data matrix. Phylogenetic analysis supported the monophyly of the C. ignita species group and divided its species into well-supported clades. The resultant phylogeny was only partly in accordance with published subgroupings based on morphology. Our results suggest that several taxa currently treated as subspecies or names treated as synonyms may in fact constitute separate species. Our study provides a solid basis for further systematic investigations of this enigmatic insect group.
Containing more than a hundred species, the Chrysis ignita species group is the largest and one o... more Containing more than a hundred species, the Chrysis ignita species group is the largest and one of the most taxonomically challenging groups in its genus. It has not been possible to resolve the taxonomy of the group using traditional methods due to the lack of robust diagnostic morphological characters. Here we present the results of a molecular analysis designed to delimit species in the Chrysis ignita group for the first time; using mitochondrial sequence data for 364 in-group specimens consisting of all 18 species known to occur in Northern Europe. Two mitochondrial loci were analysed: a COI gene fragment, and a continuous DNA sequence consisting of 16S rRNA, tRNA Val , 12S rRNA and ND4. Two approaches were employed for delimiting species: (1) genetic distance analysis based on the standard COI barcode sequences and; (2) phylogenetic analysis of the COI fragment together with rRNA genes. Both analyses yielded trees with similar topology, but support values for nodes were higher using the second approach. Fifteen species were distinguished in all analyses: and C. vanlithi Linsenmaier, 1959. The specific status of C. mediata Linsenmaier, 1951 and C. solida Haupt, 1957 was not resolved. Included unidentified specimens grouped in three clusters, two of which are distinctly delimited and apparently represent cryptic species. The specific status of the unidentified samples in the third cluster remained unclear. Moreover, our data suggest the existence of additional cryptic species currently lumped under the names C. pseudobrevitarsis C. schencki Linsenmaier, 1968. In conclusion, our results derived from analysis of mitochondrial loci strongly support the specific status of the majority of currently recognised species in the Chrysis ignita species group, and suggest the existence of additional cryptic species in Northern Europe. Thus, considering the difficulties that often arise during species determination based on morphological characters, the mtDNA loci used here appear highly suitable for assisting species delimitation in this group as well as identification of specimens.
The grey wolf (Canis lupus) is an iconic large carnivore that has increasingly been recognized as... more The grey wolf (Canis lupus) is an iconic large carnivore that has increasingly been recognized as an apex predator with intrinsic value and a keystone species. However, wolves have also long represented a primary source of human¿carnivore conflict, which has led to long-term persecution of wolves, resulting in a significant decrease in their numbers, genetic diversity and gene flow between populations. For more effective protection and management of wolf populations in Europe, robust scientific evidence is crucial. This review serves as an analytical summary of the main findings from wolf population genetic studies in Europe, covering major studies from the `pre-genomic era¿ and the first insights of the `genomics era¿. We analyse, summarize and discuss findings derived from analyses of three compartments of the mammalian genome with different inheritance modes: maternal (mitochondrial DNA), paternal (Y chromosome) and biparental [autosomal microsatellites and single nucleotide polymorphisms (SNPs)]. To describe large-scale trends and patterns of genetic variation in European wolf populations, we conducted a meta-analysis based on the results of previous microsatellite studies and also included new data, covering all 19 European countries for which wolf genetic information is available: Norway, Sweden, Finland, Estonia, Latvia, Lithuania, Poland, Czech Republic, Slovakia, Germany, Belarus, Russia, Italy, Croatia, Bulgaria, Bosnia and Herzegovina, Greece, Spain and Portugal. We compared different indices of genetic diversity in wolf populations and found a significant spatial trend in heterozygosity across Europe from south-west (lowest genetic diversity) to north-east (highest). The range of spatial autocorrelation calculated on the basis of three characteristics of genetic diversity was 650-850 km, suggesting that the genetic diversity of a given wolf population can be influenced by populations up to 850 km away. As an important outcome of this synthesis, we discuss the most pressing issues threatening wolf populations in Europe, highlight important gaps in current knowledge, suggest solutions to overcome these limitations, and provide recommendations for science-based wolf conservation and management at regional and Europe-wide scales.
The ignita species group within the genus Chrysis includes over 100 cuckoo wasp species, which al... more The ignita species group within the genus Chrysis includes over 100 cuckoo wasp species, which all lead a parasitic lifestyle and exhibit very similar morphology. The lack of robust, diagnostic morphological characters has hindered phylogenetic reconstructions and contributed to frequent misidentification and inconsistent interpretations of species in this group. Therefore, molecular phylogenetic analysis is the most suitable approach for resolving the phylogeny and taxonomy of this group. We present a wellresolved phylogeny of the Chrysis ignita species group based on mitochondrial sequence data from 41 ingroup and six outgroup taxa. Although our emphasis was on European taxa, we included samples from most of the distribution range of the C. ignita species group to test for monophyly. We used a continuous mitochondrial DNA sequence consisting of 16S rRNA, tRNA Val , 12S rRNA and ND4. The location of the ND4 gene at the 3 0 end of this continuous sequence, following 12S rRNA, represents a novel mitochondrial gene arrangement for insects. Due to difficulties in aligning rRNA genes, two different Bayesian approaches were employed to reconstruct phylogeny: (1) using a reduced data matrix including only those positions that could be aligned with confidence; or (2) using the full sequence dataset while estimating alignment and phylogeny simultaneously. In addition maximum-parsimony and maximum-likelihood analyses were performed to test the robustness of the Bayesian approaches. Although all approaches yielded trees with similar topology, considerably more nodes were resolved with analyses using the full data matrix. Phylogenetic analysis supported the monophyly of the C. ignita species group and divided its species into well-supported clades. The resultant phylogeny was only partly in accordance with published subgroupings based on morphology. Our results suggest that several taxa currently treated as subspecies or names treated as synonyms may in fact constitute separate species. Our study provides a solid basis for further systematic investigations of this enigmatic insect group.
The extent of gene flow during the range expansion of non-native species influences the amount of... more The extent of gene flow during the range expansion of non-native species influences the amount of genetic diversity retained in expanding populations. Here, we analyse the population genetic structure of the raccoon dog (Nyctereutes procyonoides) in north-eastern and central Europe. This invasive species is of management concern because it is highly susceptible to fox rabies and an important secondary host of the virus. We hypothesized that the large number of introduced animals and the species' dispersal capabilities led to high population connectivity and maintenance of genetic diversity throughout the invaded range. We genotyped 332 tissue samples from seven European countries using 16 microsatellite loci. Different algorithms identified three genetic clusters corresponding to Finland, Denmark and a large 'central' population that reached from introduction areas in western Russia to northern Germany. Cluster assignments provided evidence of long-distance dispersal. Th...
Concordant differences in morphology, phenology and RAMS markers, as well as in sequenced mtDNA (... more Concordant differences in morphology, phenology and RAMS markers, as well as in sequenced mtDNA (COI, COII, cytb) and nuclear DNA (ITS2) fragments, indicate that Dolerus asper Zaddach, 1859 and Dolerus brevicornis Zaddach, 1859 are valid species. On the basis of morphology, molecular markers, and distributional records, both species are distinct from Dolerus gibbosus Hartig, 1837 (= Dolerus planatus Hartig, 1837).
... that the placement of Larentiinae may have been an artefact resulting from insufficient taxon... more ... that the placement of Larentiinae may have been an artefact resulting from insufficient taxon and character ... & Kaila (2004), as the analysis inferred Cosymbiini as the basal tribe of the ... First, feeding on Polygonaceae in the larval stage is shared by Lythriini, Rhodometrini and ...
We analyzed phylogenetic relationships of brown bear (Ursus arctos) maternal lineages in Finland ... more We analyzed phylogenetic relationships of brown bear (Ursus arctos) maternal lineages in Finland using nucleotide sequences of the mitochondrial control region of 135 individuals. A total of 14 haplotypes, which belong to 2 haplogroups, were characterized. Haplogroup I is spread across the entire bear range, whereas haplogroup D is restricted to the southeastern part of the country. Most unconventional was the concentration of haplotype C in a small territory in central Finland, which is explained by the natal philopatry of bears that descended from one or more translocated females. Analysis of Finnish brown bear haplotypes together with 15 European eastern lineage haplotypes revealed that the Romanian-Slovakian region was probably an important refuge (or route) for recolonization of Finland by brown bears after the last ice age.
... Sequences were resolved using an ABI PRISM 377 automated sequencer (Applied Biosystems, Foste... more ... Sequences were resolved using an ABI PRISM 377 automated sequencer (Applied Biosystems, Foster City, USA). ... The hypothesis of hybridization between T. griseata and T. comae receives further support from the discussion of Pittaway (1993) for closely related sphingid ...
Concordant differences in morphology, phenology and RAMS markers, as well as in sequenced mtDNA (... more Concordant differences in morphology, phenology and RAMS markers, as well as in sequenced mtDNA (COI, COII, cytb) and nuclear DNA (ITS2) fragments, indicate that Dolerus asper Zaddach, 1859 and Dolerus brevicornis Zaddach, 1859 are valid species. On the basis of morphology, molecular markers, and distributional records, both species are distinct from Dolerus gibbosus Hartig, 1837 (= Dolerus planatus Hartig, 1837). Taxonomy of the species is clarified and the neotypes of Dolerus asper Zaddach, 1859 and Dolerus brevicornis Zaddach, 1859 are designated. The synonymies of Dolerus asper Zaddach, 1859, to Dolerus planatus Hartig, 1837 and Dolerus derzavini Malaise, 1931, spec. rev. to D. asper Zaddach, 1859 are abandoned. Dolerus carbonarius Zaddach, 1859 and Dolerus fumosus Zaddach, 1859 are considered to be species inquirendae. Phylogenetic analyses of the ITS2 fragment and fragments of ITS2 + COI and ITS2 + COII yielded the topology [D. asper, (D. brevicornis, D. gibbosus)], while those of all other markers and their combinations resulted in the topology [D. brevicornis, (D. asper, D. gibbosus)]. In the latter hypothesis the clade asper + gibbosus is also supported by structural synapomorphies.
Escherichia coli ribosomes with an A to U transversion at nucleotide 1067 of their 23 S rRNA are ... more Escherichia coli ribosomes with an A to U transversion at nucleotide 1067 of their 23 S rRNA are impaired in their effective association rate constants (kcat/KM) for both EF-Tu and EF-G binding. In addition, the times that EF-G and EF-Tu spend on the ribosome during elongation are significantly increased by the A to U transversion. The U1067 mutation impairs EF-Tu function more than EF-G function. The increase in the time that EF-Tu remains bound to ribosome is caused, both by a slower rate of GTP-hydrolysis in ternary complex and by a slower EF-Tu·GDP release from the mutated ribosomes. There is, at the same time, no change in ribosomal accuracy for aminoacyl-tRNA recognition. With support from these new data we propose that nucleotide 1067 is part of the ribosomal A-site where it directly interacts with both EF-G and EF-Tu.
The raccoon dog (Nyctereutes procyonoides) is a canid that is indigenous in East Asia and alien i... more The raccoon dog (Nyctereutes procyonoides) is a canid that is indigenous in East Asia and alien in Europe, where it was introduced more than half a century ago. The aim of this study was to compare the parasite faunas associated with raccoon dogs in their native and introduced ranges, and to identify zoonotic parasite species. We examined 255 carcasses of hunted raccoon dogs from Estonia and recorded a total of 17 helminth species: 4 trematodes, 4 cestodes and 9 nematodes. The most prevalent parasite species were Uncinaria stenocephala (97.6%) and Alaria alata (68.3%). Average parasite species richness was 2.86 (the highest was 9) and only two animals were not parasitized at all. Although the infection intensity was determined by weight and not by sex, all animals infected with more than five helminth species were males. We also found that animals infected with higher numbers of helminth species fed significantly more on natural plants. Intentional consumption of grass may represent a self-medicating behaviour among raccoon dogs. We included the Estonian data into a wider comparison of raccoon dog parasite faunas and found a total of 54 helminth taxa, including 28 of zoonotic potential. In Europe, raccoon dogs are infected with a minimum of 32 helminth species of which 19 are zoonotic; in the native range they are infected with 26 species of which 17 are zoonotic. Most species were nematodes or trematodes, with fewer cestodes described. The recent increase in the number and range of raccoon dogs in Europe and the relatively high number of zoonotic parasite taxa that it harbours suggests that this species should be considered an important source of environmental contamination with zoonotic agents in Europe.
In the Baltic countries, the two zoonotic diseases, alveolar echinococcosis (AE) caused by Echino... more In the Baltic countries, the two zoonotic diseases, alveolar echinococcosis (AE) caused by Echinococcus multilocularis, and cystic echinococcosis (CE) caused by Echinococcus granulosus, are of increasing public health concern. Observations from Estonia, Latvia and Lithuania indicate that the distribution of both parasites is wider in the Baltics than previously expected. In this paper, we review and discuss the available data, regarding both parasitoses in animals and humans, from the Baltic countries and selected adjacent regions. The data are not easily comparable but reveal a worrisome situation as the number of human AE and CE cases is increasing. Despite improvements in diagnostics and treatment, AE has a high morbidity and mortality in the Baltic region. For the control of both zoonoses, monitoring transmission patterns and timely diagnosis in humans as well as the development of local control programs present major challenges.
Alveolar echinococcosis, caused by the tapeworm Echinococcus multilocularis, is one of the most s... more Alveolar echinococcosis, caused by the tapeworm Echinococcus multilocularis, is one of the most severe parasitic diseases in humans and represents one of the 17 neglected diseases prioritised by the World Health Organisation (WHO) in 2012. Considering the major medical and veterinary importance of this parasite, the phylogeny of the genus Echinococcus is of considerable importance; yet, despite numerous efforts with both mitochondrial and nuclear data, it has remained unresolved. The genus is clearly complex, and this is one of the reasons for the incomplete understanding of its taxonomy. Although taxonomic studies have recognised E. multilocularis as a separate entity from the Echinococcus granulosus complex and other members of the genus, it would be premature to draw firm conclusions about the taxonomy of the genus before the phylogeny of the whole genus is fully resolved. The recent sequencing of E. multilocularis and E. granulosus genomes opens new possibilities for performing in-depth phylogenetic analyses. In addition, whole genome data provide the possibility of inferring phylogenies based on a large number of functional genes, i.e. genes that trace the evolutionary history of adaptation in E. multilocularis and other members of the genus. Moreover, genomic data open new avenues for studying the molecular epidemiology of E. multilocularis: genotyping studies with larger panels of genetic markers allow the genetic diversity and spatial dynamics of parasites to be evaluated with greater precision. There is an urgent need for international coordination of genotyping of E. multilocularis isolates from animals and human patients. This could be fundamental for a better understanding of the transmission of alveolar echinococcosis and for designing efficient healthcare strategies.
Samples were collected and stored at 80°C for 1 week to avoid the risk of infection with any Ec... more Samples were collected and stored at 80°C for 1 week to avoid the risk of infection with any Echinococcus spp. eggs present. Approximately 250 mg was placed into 2-mL tubes, heated at 65°C for 15 min, and stored at 80°C. The heating and cooling procedure helps to break the parasite egg shells, enabling more efficient DNA extraction. DNA was extracted by using the QIAamp DNA Stool Mini Kit (QIAGEN, Hilden, Germany) according to the manufacturer's instructions. were designed to amplify short sequences of mitochondrial DNA. The Echinococcus multilocularis tapeworm-specific primer pair (EMfor1 and EMrev1) amplifies tRNA-Ile/Lys, the E. granulosus tapeworm-specific (EGfor1 and EGrev1) and fox-specific (F1 and RVu) primer pairs amplify 12S rRNA, and the dog-specific (Dog1f and HW1r) primer pair amplifies part of the mitochondrial DNA control region. Although the 3 primer pairs were highly specific, the dog primers can potentially amplify gray wolf (Canis lupus) DNA. In other similar studies, only the parasite (1) or the host species
The raccoon dog (Nyctereutes procyonoides) is an alien species in Europe and an important vector ... more The raccoon dog (Nyctereutes procyonoides) is an alien species in Europe and an important vector of zoonotic diseases. However, compared to the red fox (Vulpes vulpes), less attention has been paid to the raccoon dog as a potentially important host for Echinococcus multilocularis, the infective agent of alveolar echinococcosis, which is an emerging infectious disease with a high mortality rate. We examined the small intestines of 249 Estonian raccoon dogs and found 1.6% of individuals to be infected with E. multilocularis. The relatively large difference between this prevalence and that found in sympatric red foxes (31.5%) sampled during the same time period might be due to differences in diet: red foxes consume significantly more arvicolid rodents - the main intermediate hosts of the parasite - especially during the coldest period of the year when raccoon dogs hibernate. Nonetheless, given the relatively high density of raccoon dogs, our results suggest that the species also represents an important definitive host species for E. multilocularis in Estonia. Compared with other countries in Europe where E. multilocularis-infected raccoon dogs have been recorded (Latvia, Lithuania, Poland, Germany, and Slovakia), the prevalence in Estonia is low. The longer hibernation period of raccoon dogs at higher latitudes may explain this pattern. Both mitochondrial and nuclear loci were analysed for Estonian isolates: based on EmsB microsatellite genotyping the Estonian isolates shared an identical genotype with E. multilocularis in northern Poland, suggesting a common history with this region. The data from more than a quarter of the mitochondrial genome (3558 bp) revealed two novel haplotypes specific to Estonia and placed them into the same haplogroup with isolates from other European regions. Considering that the raccoon dog is becoming increasingly widespread and is already relatively abundant in several countries in Europe, the role of the species must be taken into account when assessing the E. multilocularis related risks to public health.
Background: Echinococcus granulosus and E. multilocularis are tapeworm parasites of major medical... more Background: Echinococcus granulosus and E. multilocularis are tapeworm parasites of major medical and veterinary importance, causing cystic and alveolar echinococcosis, respectively. Both diseases are listed among the most severe parasitic diseases in humans, representing 2 of the 17 neglected diseases prioritised by the World Health Organisation. However, little is known about the role of urban animals in transmission of both parasite species.
Previous experiments have shown that the top of helix 90 of 23S rRNA is highly important for the ... more Previous experiments have shown that the top of helix 90 of 23S rRNA is highly important for the ribosomal peptidyltransferase activity and might be part of the donor (P) site. Developing on these studies, mutations in the 23S rRNA at the highly conserved positions G2505, G2582, and G2583 were investigated. None of the mutations affected assembly, subunit association, or the capacity of tRNA binding to A and P sites. A "selective transpeptidation assay" revealed that the mutations specifically impaired peptide bond formation. Results with a modified "fragment" assay using the minimal donor substrate pA-fMet are consistent with a model where the nucleotides psiGG2582 form a binding pocket for C75 of the tRNA.
The ignita species group within the genus Chrysis includes over 100 cuckoo wasp species, which al... more The ignita species group within the genus Chrysis includes over 100 cuckoo wasp species, which all lead a parasitic lifestyle and exhibit very similar morphology. The lack of robust, diagnostic morphological characters has hindered phylogenetic reconstructions and contributed to frequent misidentification and inconsistent interpretations of species in this group. Therefore, molecular phylogenetic analysis is the most suitable approach for resolving the phylogeny and taxonomy of this group. We present a wellresolved phylogeny of the Chrysis ignita species group based on mitochondrial sequence data from 41 ingroup and six outgroup taxa. Although our emphasis was on European taxa, we included samples from most of the distribution range of the C. ignita species group to test for monophyly. We used a continuous mitochondrial DNA sequence consisting of 16S rRNA, tRNA Val , 12S rRNA and ND4. The location of the ND4 gene at the 3 0 end of this continuous sequence, following 12S rRNA, represents a novel mitochondrial gene arrangement for insects. Due to difficulties in aligning rRNA genes, two different Bayesian approaches were employed to reconstruct phylogeny: (1) using a reduced data matrix including only those positions that could be aligned with confidence; or (2) using the full sequence dataset while estimating alignment and phylogeny simultaneously. In addition maximum-parsimony and maximum-likelihood analyses were performed to test the robustness of the Bayesian approaches. Although all approaches yielded trees with similar topology, considerably more nodes were resolved with analyses using the full data matrix. Phylogenetic analysis supported the monophyly of the C. ignita species group and divided its species into well-supported clades. The resultant phylogeny was only partly in accordance with published subgroupings based on morphology. Our results suggest that several taxa currently treated as subspecies or names treated as synonyms may in fact constitute separate species. Our study provides a solid basis for further systematic investigations of this enigmatic insect group.
Containing more than a hundred species, the Chrysis ignita species group is the largest and one o... more Containing more than a hundred species, the Chrysis ignita species group is the largest and one of the most taxonomically challenging groups in its genus. It has not been possible to resolve the taxonomy of the group using traditional methods due to the lack of robust diagnostic morphological characters. Here we present the results of a molecular analysis designed to delimit species in the Chrysis ignita group for the first time; using mitochondrial sequence data for 364 in-group specimens consisting of all 18 species known to occur in Northern Europe. Two mitochondrial loci were analysed: a COI gene fragment, and a continuous DNA sequence consisting of 16S rRNA, tRNA Val , 12S rRNA and ND4. Two approaches were employed for delimiting species: (1) genetic distance analysis based on the standard COI barcode sequences and; (2) phylogenetic analysis of the COI fragment together with rRNA genes. Both analyses yielded trees with similar topology, but support values for nodes were higher using the second approach. Fifteen species were distinguished in all analyses: and C. vanlithi Linsenmaier, 1959. The specific status of C. mediata Linsenmaier, 1951 and C. solida Haupt, 1957 was not resolved. Included unidentified specimens grouped in three clusters, two of which are distinctly delimited and apparently represent cryptic species. The specific status of the unidentified samples in the third cluster remained unclear. Moreover, our data suggest the existence of additional cryptic species currently lumped under the names C. pseudobrevitarsis C. schencki Linsenmaier, 1968. In conclusion, our results derived from analysis of mitochondrial loci strongly support the specific status of the majority of currently recognised species in the Chrysis ignita species group, and suggest the existence of additional cryptic species in Northern Europe. Thus, considering the difficulties that often arise during species determination based on morphological characters, the mtDNA loci used here appear highly suitable for assisting species delimitation in this group as well as identification of specimens.
In Estonia, both genetic (maternal lineages) and osteological data (consumption patterns) indicat... more In Estonia, both genetic (maternal lineages) and osteological data (consumption patterns) indicate that sheep husbandry has been consistently a prime source of subsistence for societies in the past. Today, sheep farming in Estonia is rather small-scale and economically of minor importance. However, there is growing interest in native sheep and their products - meat, wool and skin. This trend has provoked questions about the origin of the local native sheep and their connections with ancient ovine populations. One method we use to approach this issue is the presence of endogenous retroviruses in the genomes of ancient and modern sheep from north-eastern Europe, potentially showing us whether the studied individuals have affinities with the “primitive” northern European breeds and/or with the modern breeds. Another approach we use to assess the relations and development of sheep populations is morphometry. Here the osteometric data from archaeological sheep remains from the Late Bronze Age onwards in Estonia are compared to those of modern European breeds.
The maternal lineages of Estonian ancient sheep extend back in time for almost 3000 years, with a... more The maternal lineages of Estonian ancient sheep extend back in time for almost 3000 years, with affinities to modern breeds, including the native Kihnu (an island in the Baltic) sheep. Together with osteological data these results prove that sheep husbandry was constant and one of the main sources of subsistence for societies in the past. In Estonia today, sheep farming is rather small-scale and economically of minor importance. However, there is growing interest in native sheep and their products like meat, wool and skin. This trend has provoked questions about the origin of the Kihnu native sheep and their connections with other populations of ancient sheep. One method we use to approach this issue is the presence of endogenous retroviruses in the sheep genome, related to the pathogenic Jaagsiekte sheep retrovirus. Past studies have proved retrovirus integrations to be a useful marker for the identification of “primitive” populations and can therefore show whether the Kihnu sheep have affinities with the old northern European and/or modern breeds. The same retrotypes are also tested within the ancient specimens from the Late Bronze Age to the Modern Period from Estonia and other north-eastern European regions. Additionally, phylogenetic networks of mitochondrial sequences help to position the Estonian ancient and modern sheep within a wider spatial and temporal context. Another approach we use to assess the relations and development of sheep populations is linear morphometrics. Here we present the osteometric data for archaeological sheep specimens from the Late Bronze Age onwards in Estonia and compare them to modern populations of various breeds.
Sheep (Ovis aries) were among the first domesticated animals to appear in Estonia in the late Neo... more Sheep (Ovis aries) were among the first domesticated animals to appear in Estonia in the late Neolithic around 3000 BC and became one of the most widespread livestock species from the Late Bronze Age onwards. To study genetic diversity and temporal changes in sheep populations in Estonia and to compare them with other ancient and modern sheep in Eurasia, fragments of the mtDNA D-loop and Y-chromosome SRY gene of ancient as well as of modern native sheep have been analysed. The results have revealed: (1) two lineages, A and B; (2) SNP G-oY1 of the Y-chromosome; (3) high genetic diversity through time; (4) possible population expansion during the first centuries of the Middle Ages; (5) continuity of some of the haplotypes from the Late Bronze Age to present-day; (6) differences in genetic diversity between ancient and modern populations. The results coincide with the known history of sheep, which is based on zooarchaeological and written records. It seems that despite the observed fluctuations in the ancient sheep populations and changes in ecology, power regime, and other influential historical events, the matrilineal basis of sheep has remained largely the same, and has continued to modern native sheep. In future research phenotypic, whole nuclear and more comprehensive Y-chromosome analyses are required. Pilot NGS data based on 30 sheep samples from Estonia, Russia and Latvia have shown promising endogenous DNA content (... to 80%). Also continuous work with osteological material is planned to shed more light on past livestock exploitation and animal husbandry.
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Papers by Urmas Saarma