Citizen science and early detection of invasive species: phenology of first occurrences of <Emphasis Type="Italic">Halyomorpha halys</Emphasis> in Southern Europe

Early detection of invasive alien species and the ability to track their spread are critical for undertaking appropriate management decisions. Citizen science surveys are potentially valuable tools for quickly obtaining information on biodiversity and species distributions. The Asian brown marmorated stink bug, Halyomorpha halys, is an invasive pest of agricultural crops and a dwelling nuisance. Halyomorpha halys was first recorded in Italy in 2012 in Emilia Romagna, one of the most important fruit producing regions of Europe. To rapidly obtain data on its distribution in the newly invaded area, a survey that combined citizen science and active search was set up using multimedia channels. Data concerning when, where and how the bugs were spotted were collected, together with photographs and specimens. The survey detected established breeding populations in different areas of Northern Italy and Southern Switzerland, indicating a potentially high risk for crop damage that extends beyond the territories of first detection. Furthermore, new data on H. halys phenology, host plants, voltinism and behaviour were obtained. The importance of citizen science in early detection of introduced pest species is highlighted. This paper also provides a picture-based key to recognize H. halys from similar pentatomids in the world.     

Predation of <Emphasis Type="Italic">Halyomorpha halys</Emphasis> (Hemiptera: Pentatomidae) from Web-Building Spiders Associated with Anthropogenic Dwellings

The brown marmorated stink bug, or Halyomorpha halys, is an invasive pest in North America and Europe that causes severe agricultural damage and nuisance problems for homeowners; and it is originally from China, Taiwan, and the Republic of Korea. While the natural enemy community of H. halys has been evaluated in several agroecosystems, it has not been examined where H. halys overwinters in anthropogenic structures. The aims of the current study were to evaluate 1) whether spider webs commonly found in the home and yard can successfully ensnare H. halys, 2) whether entanglement resulted in consumption by spiders inhabiting the webs, and 3) how frequently H. halys becomes entangled in webs under ambient conditions. To accomplish this, adult H. halys were introduced into webs in and near anthropogenic structures in West Virginia and Maryland, United States, and the behavior of spiders was observed for 5-min intervals at 0, 1, 2, and 24 h after introduction. In addition, a survey of webs was performed to determine the frequency with which spiders naturally capture H. halys inside buildings and in the landscape. Overall, the study found seven spider families in anthropogenic structures. Adult H. halys that were introduced into the webs of Theridiidae, Pholcidae, or Agelenidae had a greater than 50% chance of being ensnared and consumed. Adult H. halys were found naturally most often in webs of Theridiidae. Webs with a funnel or cob web architecture had the greatest probability of ensnaring H. halys, while those with orb structures resulted in the fewest caught. In the wild, 13–20% of spider webs contained dead H. halys. Our results suggest that spiders may be an important contributing factor for mortality of H. halys at overwintering sites, and spiders in or outside homes may help reduce nuisance problems caused by H. halys.     

Invasive brown marmorated stink bug <Emphasis Type="Italic">Halyomorpha halys</Emphasis> (Stål) (Heteroptera: Pentatomidae) in Russia,Abkhazia, and Serbia: history of invasion,range expansion,early stages of establishment,and first records of damage to local crops

Halyomorpha halys is native to Asia, but was recently accidentally introduced to North America and Europe. This species was recorded for the first time in Russia in 2013 (2014) and in both Serbia and Abkhazia in 2015. In 2015–2017, we conducted surveys in Russia, Serbia, and Abkhazia and found that H. halys had not only successfully survived its first winters in new regions, but also started to establish locally, spread, increase populations, and damage local crops. In Sochi (Russia) and Abkhazia, H. halys was recorded to feed on 32 species and cultivars from 16 plants families. In Serbia, it has so far been observed on only four species from four plant families. H. halys is already widely spread in Abkhazia and causes serious damage to many crops: in 2016, the yield of peach, mandarin orange, persimmon and other crops fell down by 13.2–87.4% if compared to the long-term means of yield. The losses are likely to be mostly caused by feeding of H. halys. We conclude that H. halys was accidentally introduced to Russia in 2012–2013 most likely with woody plants imported to Sochi from Italy or Greece for urban landscaping before the 2014 Winter Olympics. The species established in Sochi and quickly moved to Krasnodar (Russia), Abkhazia, and beyond—to Georgia. Populations in Serbia most likely arrived in 2015 traveling on railway cars or in commodities on the Bucharest–Belgrade railway line. We discuss phenology and prospects of further expansion of H. halys in Europe.     

Review of the occurrence of <Emphasis Type="Italic">Halyomorpha halys</Emphasis> (Hemiptera: Heteroptera: Pentatomidae) in Italy,with an update of its European and World distribution

The regions in which Halyomorpha halys (Stål, 1855) occurs in Italy are summarized and ordered chronologically according to the date of the first record. Details and verification about the first Italian record from Liguria (often overlooked), that dates back to 2007 the introduction of H. halys in this country, are also given. First exact localities are listed for the Italian regions of Tuscany, Marche, Latium, Abruzzo, Campania, and Apulia, in which only general occurrence was known so far. We included also additional records from Trentino-Alto Adige, where the presence of H. halys has only recently been reported through a few data. Furthermore first records for Sweden, Central Macedonia (Greece), and Manitoba (Canada) are reported, as well as first intercepted specimens from Iceland.     

First records of the brown marmorated stink bug <Emphasis Type="Italic">Halyomorpha halys</Emphasis> (Stål, 1855) (Heteroptera,Pentatomidae) in Russia,Abkhazia, and Georgia

The brown marmorated stink bug Halyomorpha halys (Stål, 1855) is recorded for the first time from Russia (Khosta and Adler districts of the city of Sochi), Abkhazia (Pitsunda), and Georgia (Khobi Municipality). Outbreaks of H. halys populations were detected in all of these regions in October 2016. More than a hundred specimens congregated for overwintering were collected in Sochi during two days. According to the personal communications of the local people, the species appeared in Sochi not later than in 2013, and in Georgia not later than in 2015. It must have been brought to Sochi from Europe (from Italy) with planting material of ornamental plants for landscaping the XXII Olympic Winter Games. Thereafter it spread to Abkhazia and Georgia. Additional steps to identification of H. halys are provided for the keys to the true bugs of the European part of the former USSR (Kerzhner and Jaczewski, 1964).     

<Emphasis Type="Italic">Drosophila</Emphasis> as models to understand the adaptive process during invasion

The last few decades have seen a growing number of species invasions globally, including many insect species. In drosophilids, there are several examples of successful invasions, i.e. Zaprionus indianus and Drosophila subobscura some decades ago, but the most recent and prominent example is the invasion of Europe and North America by the pest species, Drosophila suzukii. During the invasive process, species often encounter diverse environmental conditions that they must respond to, either through rapid genetic adaptive shifts or phenotypic plasticity, or by some combination of both. Consequently, invasive species constitute powerful models for investigating various questions related to the adaptive processes that underpin successful invasions. In this paper, we highlight how Drosophila have been and remain a valuable model group for understanding these underlying adaptive processes, and how they enable insight into key questions in invasion biology, including how quickly adaptive responses can occur when species are faced with new environmental conditions.     

Modelling the potential geographic distribution of <Emphasis Type="Italic">Trissolcus japonicus</Emphasis>: a biological control agent of the brown marmorated stink bug, <Emphasis Type="Italic">Halyomorpha halys</Emphasis>

Trissolcus japonicus Ashmead (Hymenoptera: Scelionidae) is an endoparasitoid of the eggs of the brown marmorated stink bug, Halyomorpha halys Stål (Hemiptera: Pentatomidae), a major agricultural pest native to China, Japan, South Korea and Taiwan. We used CLIMEX to estimate the potential global distribution of T. japonicus with particular reference to New Zealand. In its native range the model predicts the presence, or a potential expansion, of T. japonicus into most of humid-subtropical and humid-continental areas. Globally, the model projects that many temperate, Mediterranean and subtropical areas could suit the establishment of T. japonicus. In New Zealand, the north appears moderately to highly suitable for T. japonicus, while southern regions are mostly marginal. The risk posed by T. japonicus to non-target species in New Zealand is predicted to vary between different non-targets. CLIMEX projections of the potential distribution of T. japonicus provide guidance for release sites of this parasitoid if approved for importation and release in New Zealand.     

Control effect on the brown-marmorated stink bug, <Emphasis Type="Italic">Halyomorpha halys</Emphasis> (Hemiptera: Pentatomidae), by combined spraying of pyrethroid and neonicotinoid insecticides in apple orchards in northern Japan

Control of Halyomorpha halys (Stål) adults by combined spraying of pyrethroid and neonicotinoid insecticides has been investigated in apple orchards in northern Japan in 2008 and 2009. Insecticides were selected to control not only H. halys but also the common lepidopteran pests Carposina sasakii Matsumura, Adoxophyes orana fasciata Walsingham, and Phyllonorycter ringoniella Matsumura. Dinotefuran was sprayed in mid-May and early July, fenpropathrin in late June, and bifenthrin in late July. H. halys adults were released into apple trees at 3–5-day intervals from mid-May to mid-August. In this spraying program, survival of the adults increased 6 days after spraying, but relatively few injury marks were noted throughout the growing season. On the other hand, in the conventional spraying program, the survival ratio of the adults increased rapidly 3 days after spraying—especially after the organophosphate applied in mid-May, late June, and late July—and increases in the number of injury marks synchronized well with survival in both years. This spray program reduced the number of fruit injury marks to one-fifth that of the conventional spraying program and one-tenth that of fungicides alone.     

A new invasive species of buprestid beetles in the Russian fauna: <Emphasis Type="Italic">Lamprodila</Emphasis> (<Emphasis Type="Italic">Palmar</Emphasis>) <Emphasis Type="Italic">festiva</Emphasis> (L.) (Coleoptera,Buprestidae), a pest of Cupressaceae

The buprestid beetle Lamprodila (Palmar) festiva (L.) (Coleoptera, Buprestidae, Poecilonoini), an invasive pest of cultivated Cupressaceae such as Thuja, Chamaecyparis, Cupressus, and Juniperus in the Greater Sochi area (the Russian Black Sea coast), is recorded for the first time for Russia and the former USSR. Data on the systematic position, diagnostic characters, distribution, host plants, biology of the new pest, and its damage to urban plantations are discussed. Recent records of this species in the new areas of Central and Eastern Europe as well as on the Black Sea coast of the Caucasus suggest expansion of its range during the recent decades.     

Cryptic invasion of Italian pool frogs (<Emphasis Type="Italic">Pelophylax bergeri</Emphasis>) across Western Europe unraveled by multilocus phylogeography

Human introductions of exotic amphibians can have catastrophic effects on native species. However, they usually remain unnoticed without genetic tools when species are difficult to distinguish morphologically. In Western Europe, pool frogs (Pelophylax sp.) make a worrisome case: recent genetic data showed the presence of Italian (Pelophylax bergeri) mtDNA haplotypes within the range of the threatened European Pelophylax lessonae, two morphologically similar taxa. Here we conduct a multilocus phylogeographic and population genetic survey of European and Italian pool frogs (combining present and historic samples), to investigate the origin(s) and consequences of potential introductions. Results are unequivocal: we show that the alien P. bergeri have extensively invaded France and north-Alpine Switzerland, and have also deeply introgressed with P. lessonae, which has led to the complete replacement of most populations. Alien specimens have probably been translocated multiple times from Central Italy at least prior to the 1960s. Based on our dense sampling, only two areas, north and south of the Alps still host native pool frogs in Switzerland, the Joux Valley near the French border and the canton of Ticino, respectively. Importantly, we show that these last P. lessonae populations are remnants of a private genetic diversity specific to Western Europe, which vanished during the P. bergeri’s invasion. Our study emphasizes the risk of genetic pollution during invasion by human-introduced taxa and brings alarming concern regarding uncontrolled amphibian translocations. Moreover, it demonstrates the necessity for genetic surveys to detect and monitor these invasions, especially where species determination is problematic.     

Geographical Ecocline Polymorphism of the Distribution of Mitochondrial Haplotypes of <Emphasis Type="Italic">Adalia bipunctata</Emphasis> Linnaeus, 1758 (Coleoptera: Coccinellidae) of Norway

Adalia bipunctata Linnaeus, 1758 is a classical object for studying polymorphism of natural populations. Application of genetic markers significantly increased the possibilities of analysis of natural populations of ladybirds and opened the possibility of studying ecological adaptations at the genetic level. We analyze the geographical distribution of the mitochondrial haplotypes of A. bipunctata, determined on the basis of the variability of the mitochondrial cox1 gene in the natural populations of Norway. Two new mitochondrial haplotypes of A. bipunctata are described. We observe a geographical cline in the haplotypes frequencies from south to north. The main haplotype H1 of the central Europe is gradually replaced in the north by the haplotype H7. A possible reason for the observed geographical cline is a selective advantage of beetles with haplotype H7 in areas with a northern coastal climate.     

Identifying the ecological and societal consequences of a decline in <Emphasis Type="Italic">Buxus</Emphasis> forests in Europe and the Caucasus

The potential impact of new invasive tree pests and diseases is usually quantified in economic terms. The ecological and social impacts are less often assessed. Using a comprehensive literature review we assess the potential ecological and social impact of two non-native invasive species (the box tree moth, Cydalima perspectalis and the fungus Calonectria pseudonaviculata) that threaten the survival of box tree, Buxus spp. in forests in Europe and the Caucasus. A total of 132 fungi, 12 chromista (algae), 98 invertebrate and 44 lichens were found to use Buxus spp. Of these, 43 fungi, 3 chromista and 18 invertebrate species have only been recorded on Buxus spp., suggesting that these species are obligate on Buxus spp. and are most at risk from in the loss of Buxus spp. due to these invasive pest and disease species. Buxus spp. was shown to be important for soil stability and water quality but there was no information on other ecosystem functions provided by Buxus spp. Buxus was found to be of considerable historical cultural importance but there was very limited information on current social values and uses. Buxus trees, wood and leaves are associated with different folklore and sacred rites which are still particularly important in the Caucasus. While we could not find any assessment of the economic value of Buxus forests the biodiversity, cultural and social values of Buxus identified here indicate that its loss could have major indirect and non-market economic effects. This work highlights the importance of studying the ecological and societal implications of biological invasions.     

Walnut husk fly, <Emphasis Type="Italic">Rhagoletis completa</Emphasis> (Diptera: Tephritidae), invades Europe: invasion potential and control strategies

Rhagoletis completa Cresson (Diptera: Tephritidae) is native to North America and invaded Western Europe in the late 1980s, causing important damage to its principal host, walnut (Juglans spp.). In this review, we summarize the important elements of R. completa’s biology, phytosanitary status and methods used in Europe for its control, and then present the main conclusions associated with a completed risk analysis performed in 2014 to evaluate the dispersion and establishment potential of R. completa in Europe. The walnut husk fly was initially identified in Switzerland (1988) and Italy (1991), from where it spread to at least seven additional countries: France, Spain, Germany, Austria, Croatia, Slovenia and Hungary. R. completa has not reached the limits of its potential distribution. The main dissemination pathways within Europe include: (1) natural adult dissemination; (2) adult hitchhiker behaviour; and, to a lesser extent, (3) transportation of larvae in fresh fruits. R. completa host plants are widely distributed in Europe, either as isolated wild trees or in orchards, favouring the probability of fly establishment in currently fly-free areas. In addition, the European territories where Juglans species are present share biogeographic similarities. In orchards where R. completa is present and uncontrolled, 100% of walnut trees can be infested, causing losses in walnut yields of up to 80%. The negative effect is low (<10% yield loss) under phytosanitary control, although additional costs must also be considered to support specific monitoring for R. completa. The information presented here underlines a strong need for better walnut husk fly monitoring across European countries, as well as for increasing efforts to develop biological methods to control this emerging pest.     

The mitochondrial DNA of <Emphasis Type="Italic">Dictyostelium discoideum</Emphasis>: complete sequence,gene content and genome organization

We present an overview of the gene content and organization of the mitochondrial genome of Dictyostelium discoideum. The mitochondria genome consists of 55,564?bp with an A + T content of 72.6%. The identified genes include those for two ribosomal RNAs (rnl and rns), 18 tRNAs, ten subunits of the NADH dehydrogenase complex (nad1, 2, 3, 4, 4L, 5, 6, 7, 9 and 11), apocytochrome b (cytb), three subunits of the cytochrome oxidase (cox1/2 and 3), four subunits of the ATP synthase complex (atp1, 6, 8 and 9), 15 ribosomal proteins, and five other ORFs, excluding intronic ORFs. Notable features of D. discoideum mtDNA include the following. (1) All genes are encoded on the same strand of the DNA and a universal genetic code is used. (2) The cox1 gene has no termination codon and is fused to the downstream cox2 gene. The 13 genes for ribosomal proteins and four ORF genes form a cluster 15.4?kb long with several gene overlaps. (3) The number of tRNAs encoded in the genome is not sufficient to support the synthesis of mitochondrial protein. (4) In total, five group I introns reside in rnl and cox1/2, and three of those in cox1/2 contain four free-standing ORFs. We compare the genome to other sequenced mitochondrial genomes, particularly that of Acanthamoeba castellanii.     

A PCR-based analysis of plant DNA reveals the feeding preferences of <Emphasis Type="Italic">Apolygus lucorum</Emphasis> (Heteroptera: Miridae)

The mirid bug Apolygus lucorum (Meyer-Dür) (Heteroptera: Miridae) is a severe pest of cotton and other crops in China. The feeding preferences of this pest are unclear due to its frequent movement among different host plants and the inconspicuous signs of its feeding. Here, we present results of a field trial that used direct observation of bug densities and a PCR-based molecular detection assay to detect plant DNA in bugs to explore relationships between A. lucorum population abundance and its feeding preference between two host plants, Humulus scandens (Loureiro) Merrill and Medicago sativa L. The field-plot samples showed that A. lucorum adults generally prefer flowering host plants. Its density was significantly higher on flowering H. scandens than on seedlings of M. sativa, and a similarly higher bug density was observed on flowering M. sativa than on seedlings of H. scandens. In the laboratory, we designed two pairs of species-specific primers targeting the trnL-F region for H. scandens and M. sativa, respectively. The detectability of plant DNA generally decreased with time post-feeding, and the half-life of plant DNA detection (DS₅₀) in the gut was estimated as 6.26 h for H. scandens and 3.79 h for M. sativa with significant differences between each other. In mirid bugs exposed to seedlings of H. scandens and flowering M. sativa, the detection rate of M. sativa DNA was significantly higher than that of H. scandens. Meanwhile, in mirid bugs exposed to seedlings of M. sativa and flowering H. scandens, a significantly higher detection rate of H. scandens DNA was found. We developed a useful tool to detect the remaining plant food species specifically from the gut of A. lucorum in the current study. We provided direct evidence of its feeding preference between H. scandens and M. sativa at different growth stages, which strongly supported a positive correlation between population abundance and feeding preference of A. lucorum on different plants under field conditions. The findings provide new insights into the understanding of A. lucorum’s feeding preference, and are helpful for developing the strategies to control this pest.     

Reaping benefits from an invasive species: role of <Emphasis Type="Italic">Harmonia axyridis</Emphasis> in natural biological control of <Emphasis Type="Italic">Aphis glycines</Emphasis> in North America

Harmonia axyridis (Pallas) (Coleoptera: Coccinellidae) is an invasive species present in numerous agroecosystems in North America. Despite adverse impacts as a threat to native biodiversity, a nuisance household invader and a pest in fruit production, H. axyridis also plays a beneficial role as a major component of assemblages of generalist predators in several agricultural crops. Here, we review the role of H. axyridis as a natural enemy of Aphis glycines Matsumura (Hemiptera: Aphididae), an invasive pest of soybean, Glycine max (L.) Merrill (Fabales: Fabaceae), in North America. Harmonia axyridis is often the most abundant predator species attacking A. glycines in soybean agroecosystems. This predator has the potential to both prevent and suppress A. glycines outbreaks. Further studies are needed to fully understand and utilize the potential of H. axyridis as a natural enemy in the management of A. glycines and other agricultural pests in agroecosystems worldwide.     

<Emphasis Type="Italic">Hexamermis popilliae</Emphasis> n. sp. (Nematoda: Mermithidae) parasitizing the Japanese beetle <Emphasis Type="Italic">Popillia japonica</Emphasis> Newman (Coleoptera: Scarabaeidae) in Italy

A new species of mermithid nematode, Hexamermis popilliae n. sp. (Nematoda: Mermithidae) is described from the Japanese beetle Popillia japonica Newman in Italy, an area of new introduction for this invasive pest. The combination of the following characters separates H. popilliae from other members of the genus Hexamermis Steiner, 1924: adult head obtuse; amphidial pouches slightly posterior to lateral head papillae in female but adjacent to lateral head papillae in males; amphidial openings large, well developed; amphidial pouches elliptical in females and oblong in males; cuticular vulvar cone well developed, vulvar lips greatly reduced or lacking, vagina curved at tip where meeting uteri, without reverse bend (not S-shaped), spicules slightly curved, with a slight bend in the basal portion, approximately equal to body width at cloaca. This is the first record of a species of Hexamermis parasitizing the Japanese beetle Popillia japonica. The only previous mention of mermithid nematodes from P. japonica was an undescribed species of Psammomermis in North America. Hexamermis popilliae will be evaluated as a potential biological control agent in an integrated control program of the Japanese beetle in Italy.     

Genetic diversity and invasion history of the European subterranean termite <Emphasis Type="Italic">Reticulitermes urbis</Emphasis> (Blattodea,Termitoidae, Rhinotermitidae)

Biological invasions are among key factors of ecological changes, and social insects appear as highly successful invasive animals. Subterranean termites of the holarctic genus Reticulitermes are present in Europe with six native and one invasive (the nearctic R. flavipes) species. The species R. urbis shows a disjunct distribution in the Western Balkans, Eastern Italy and Southern France. Previous molecular and population genetics data suggested that the taxon originated from the Balkans, and that Italian and French populations are invasive, but it is still unknown how many introduction events occurred and from which Balkan source populations. To address these questions, a population genetics analysis was performed on a larger sampling than previous studies, using mitochondrial cytochrome oxidase II and 6 microsatellite markers on 47 colonies collected across the whole distribution area. Mitochondrial analysis confirmed the presence of two major lineages where colonies from Balkans, Italy, and France intermingle. Similarly, microsatellite loci analysis indicated the presence of two genetic clusters, though not corresponding to the two mitochondrial clades, each including colonies from the three sampled areas and with individuals showing mixed cluster membership. Overall, French and Italian populations showed indications of bottleneck (reduced genetic diversity and change of allele frequencies) and do not appear genetically differentiated from the Balkan population. Results presented here support a history of multiple introductions in Italy and France, in a scenario consistent with continuous exchanges between native and invasive areas, as expected along human trades routes.     

The earliest samples of <Emphasis Type="Italic">Hymenoscyphus albidus</Emphasis> vs. <Emphasis Type="Italic">H. fraxineus</Emphasis> in Estonian mycological herbaria

In 2006, the alien and pathogenic Hymenoscyphus fraxineus was differentiated by molecular methods from the long recognized saprotrophic H. albidus . Today, H. fraxineus seems to have replaced H. albidus in several countries, but the exact year of arrival of H. fraxineus in northern Europe is still debated. Investigations of herbaria specimens might help to ascertain it. Before the epidemic, H. albidus was not of significant interest to mycologists in the Baltic area and was not eagerly sampled and deposited. Nevertheless, the TAAM herbarium in Estonia holds 13 putative H. albidus specimens that were collected 1966–2006. Using newly developed species-specific PCR primers to differentiate between H. albidus and H. fraxineus, all available herbarium samples from TAAM were identified. In addition, the primers were also tested on pure cultures and ascocarps of H. fraxineus and H. albidus, symptomatic petioles of F. excelsior, pure cultures of 10 non-target fungal species, and 10 different soil samples. With the exception of the oldest specimen in TAAM from 1966 (collected in Lithuania), all herbarium samples were identified as H. fraxineus. The first record of H. fraxineus in Estonia dates back to 1997 and apparently represents the oldest record of the species in Europe. In spite of this, symptoms based on reliable observations were not detected until 2003 in Estonia. This temporal shift is difficult to explain. Possible reasons may represent (i) overlooking of symptoms during the first years; (ii) typical lag phase at the start of an epidemic; (iii) an abrupt shift towards higher virulence of H. fraxineus; or (iv) other environmental (climatic) factors. Closer investigations are needed to disentangle these possible reasons.     

Analysis of <Emphasis Type="Italic">H63D</Emphasis> mutation in hemochromatosis (<Emphasis Type="Italic">HFE</Emphasis>) gene in populations of Central Eurasia

An analysis of the frequency of H63D (c.187C>G) mutations in the HFE gene in 19 populations from Central Eurasia demonstrated that the distribution of the mutation in the region of interest was not uniform and that there were the areas of H63D accumulation. The investigation of three polymorphic variants, c.340+4T>C (rs2071303, IVS2(+4)T>C), c.893-44T>C (rs1800708, IVS4(-44)T>C), and c.1007-47G>A (rs1572982, IVS5(-47)A>G), in the HFE gene in individuals homozygous for H63D mutations in the HFE gene revealed the linkage of H63D with three haplotypes, *CTA, *TTG, and *TTA. These findings indicated the partial spread of the mutation in Central Eurasia from Western Europe, as well as the possible repeated appearance of the mutation on the territory on interest.