Invasion biology of the Chinese mitten crab Eriochier sinensis: A brief review

The Chinese mitten crab Eriocheir sinensis is a native of freshwater and estuarine habitats along the east coast of Asia. Invasive populations have existed in northern Europe since the early 20th century, and more recently a breeding population has become established in the San Francisco Bay system along the west coast of North America. Ballast water is the most probable vector for both invasions, although there is also potential for escape from ethnic markets and from the ornamental aquarium industry. Invasive populations of mitten crabs have caused millions of dollars in economic and ecological damage. Economic impacts center largely on the burrowing activity of the crabs, which damages stream banks and levees, and the annual spawning migration, which interferes with fishing activities and irrigation projects. Chinese mitten crabs have recently appeared in the Chesapeake and Delaware Bays on the east coast of the USA, and there are confirmed reports of breeding females in both estuaries. The potential for large populations of mitten crabs in these estuaries has not been determined. This paper presents a review of the biology and ecology of native and invasive populations of the species and provides recommendations for research relevant to the prediction of future mitten crab invasions.     

Molecular connectedness between self and none self-sustainable populations of Chinese mitten crab (Eriocheir sinensis, H. Milne Edwards, 1853) with focus to the Swedish Lake Vänern and the Oder and Vistula River in Poland

The highly invasive Chinese mitten crabs (Eriocheir sinensis) was studied to reveal the genetic relationship between three none self-sustainable population within the Polish Oder and Vistula River and Lake V?nern in Sweden. Crabs from self-sustainable populations were collected from the Elbe River (Germany), San Francisco Bay (USA) and five Asian river estuaries. Both parsimony and maximum likelihood analysis of the nuclear ITS 1 and ITS 2 regions reveal a similar topological pattern. The mitochondrial cytochrome c oxidase subunit I (COI) sequence had too low resolution to be informative. The Oder River haplotypes showed high sequence similarities to both the Elbe population and/or the native spawning areas in China. The Vistula River showed haplotypic similarity to specimens from the Oder River in Poland or the Elbe River. Also high sequence similarities were observed to the specimens from the Swedish Lake V?nern indicating to an origin from the River Elbe. Minor changes in evolutionary interpretation exist depending on how gaps are treated and the analytic method. There is a complex movement involving back and forth transoceanic colonization of the Chinese mitten crab haplotypes. Invasion routes are discussed in relation to ship transfer routes and canal waterways.     

Juvenile Chinese mitten crabs (<Emphasis Type="Italic">Eriocheir sinensis</Emphasis>) in the Thames estuary: distribution,movement and possible interactions with the native crab <Emphasis Type="Italic">Carcinus maenas</Emphasis>

The Chinese mitten crab, Eriocheir sinensis is a successful invader. Whilst non-breeding adult E. sinensis have been associated with the destruction of riverbanks, little is known about the ecology of this species in its invaded areas. This is especially true of the juveniles which are a key migratory stage. Intertidal surveys along the Thames estuary indicated an increase in the population since the 1990s. Juvenile E. sinensis were abundant in the sampled upper tidal region of the Thames, refuging under boulders in the intertidal at low tide. Seasonal differences in sampled populations were observed, with a significantly lower abundance of crabs found during winter compared to summer. Mark-recaptures indicated movement in the intertidal occurring during high tide, with an influx of new crabs evident after a single tidal cycle. Endogenous rhythms were also apparent, with peaks in activity occurring corresponding with night-time high tide conditions. E. sinensis juveniles can successfully exclude similar sized native Carcinus maenas from shelters in the laboratory, regardless of which species originally inhabited the shelter. This may have implications for native estuarine Carcinus populations, which can rely on such intertidal shelters for refuges.     

The potential for mitten crab Eriocheir sinensis H. Milne Edwards, 1853 (Crustacea: Brachyura) invasion of Pacific Northwest and Alaskan Estuaries

Eriocheir sinensis H. Milne Edwards, 1853 is on the list of top 100 invaders compiled by the International Union for Conservation of Nature and Natural Resources. The recent establishment of a large Chinese mitten crab population in San Francisco Bay and the potential for introductions from California, Asia and Europe pose a significant invasion potential for estuaries and rivers from California to Alaska. This alien species would place at risk the catchment areas of the Pacific Northwest including the economic and social activities that depend upon intact aquatic systems. An analysis of ecological conditions that define the mitten crab’s native and introduced range suggests that large stable estuaries with long flushing times are necessary to sustain significant populations. Most Pacific Northwest estuaries have limited salinity intrusion, estuarine habitat and short flushing times and face a reduced risk of population establishment. Large, stable estuaries, such as the Puget Sound, may support significant populations. River-dominated estuaries, such as the Columbia River, have flushing times less than the duration of larval development and wouldn’t support populations. An application of a temperature based larval development rate to near-shore and estuary sea surface temperatures suggests that estuaries in Oregon and Washington have sufficient thermal regimes to support larval development. Most estuary systems in Alaska have limited periods where water temperatures are above the mortality threshold for the larval stages and are at a low risk for the establishment of populations. A potential sea temperature rise of two degrees Celsius would permit larval development in Alaskan estuaries, where sufficient estuarine and freshwater habitats exist.     

Chinese mitten crab <Emphasis Type="Italic">Eriocheir sinensis</Emphasis> in the Baltic Sea—a supply-side invader?

Although the Chinese mitten crab Eriocheir sinensis (H. Milne-Edwards, 1853) (Crustacea, Decapoda, Varunidae) invaded the Baltic Sea about 80 years ago, published information on its present distribution and abundance in this region is lacking. We provide here information on its Baltic-wide distribution and long-term population dynamics. The species has been found all over the coastal Baltic Sea and also in some adjacent rivers and lakes. The Chinese mitten crab appears to have increased in abundance in recent years in the northeastern part of the Baltic Sea (Gulf of Finland, Gulf of Riga, northern Baltic Proper). Higher catch rates were observed in spring (April–June) and autumn (September–November). The size variation of crabs in different samples was low (mean carapace width 6.1–6.3 cm). Despite findings of gravid females, the reproduction of the mitten crab in the central, northern and eastern Baltic region is considered unlikely due to low salinity and the individuals caught are assumed to actively migrate into the region from the species’ main European distribution area (southeastern North Sea), certainly over 1500 km migration distance. Thus, the dynamics of the North Sea population is probably regulating, at least in part, the occurrence of the Chinese mitten crab in the Baltic Sea area.     

Genetic population structure and contemporary dispersal patterns of a recent European invader, the Chinese mitten crab, Eriocheir sinensis

Genetic studies of recently established populations are challenging because the assumption of equilibrium underlying many analyses is likely to be violated. Using microsatellites, we investigated determinants of genetic structure and migration among invasive European-Chinese mitten crab populations, applying a combination of traditional population genetic analyses and nonequilibrium Bayesian methods. Consistent with their recent history, invasive populations showed much lower levels of genetic diversity than a native Chinese population, indicative of recent bottlenecks. Population differentiation was generally low but significant and especially pronounced among recently established populations. Significant differentiation among cohorts from the same geographical location (River Thames) suggests the low effective population size and associated strong genetic drift that would be anticipated from a very recent colonization. An isolation-by-distance pattern appears to be driven by an underlying correlation between geographical distance and population age, suggesting that cumulative homogenizing gene flow reduces founder bottleneck-associated genetic differentiation between longer-established populations. This hypothesis was supported by a coalescent analysis, which supported a drift + gene flow model as more likely than a model excluding gene flow. Furthermore, admixture analysis identified several recent migrants between the UK and Continental European population clusters. Admixture proportions were significantly predicted by the volume of shipping between sites, indicating that human-mediated transport remains a significant factor for dispersal of mitten crabs after the initial establishment of populations. Our study highlights the value of nonequilibrium methods for the study of invasive species, and also the importance of evaluating nonequilibrium explanations for isolation by distance patterns.     

African Origin and Europe-Mediated Global Dispersal of The Cyanobacterium Microcystis aeruginosa

Microcystis aeruginosa is a bloom-forming cyanobacteria, which currently has a cosmopolitan distribution. Since M. aeruginosa can produce toxic compounds across all continents that it inhabits, it is of major public health relevance to assess its origin and dispersal. Thus, we conducted a worldwide study using 29 isolates representative of all the main continents, and used a concatenated genetic system for phylogenetic analyses consisting of four genetic markers (spanning ca. 3,485 bp). Our results support an early origin of M. aeruginosa in the African continent, with a subsequent dispersal to establish a second genetic pool in the European continent, from where M. aeruginosa then colonized the remaining continental regions. Our findings indicate that the European population has a cosmopolitan distribution, and is genetically closer to populations from Africa and North America. Our study also highlights the utility of using a concatenated dataset for phylogenetic inferences in cyanobacteria.     

Identification and characteristics of a novel gene, <Emphasis Type="Italic">EJO1</Emphasis>, in the Chinese mitten crab (<Emphasis Type="Italic">Eriocheir japonica sinensis</Emphasis>) ovary

EJO1, a novel gene presumably involved in the ovary development of the Chinese mitten crab (Eriocheir japonica sinensis), was identified and characterized by suppression subtractive hybridization and cDNA macroarray analysis. EJO1 expression was 2.6-fold higher at stage III than at stage II during the ovary development of the mitten crab. EJO1 is 876 bp in length containing a 759 bp open reading frame which encodes a 252-amino-acid protein. Homology analysis showed that no sequence significantly matching EJO1 was found in SwissProt, so it was deduced as a novel gene (GenBank accession number: AY185917). The EJO1 protein is probably a secretion protein with a signal peptide of 17 amino acids. The pI/Mw deduced from the amino acid sequence was 6.18/28.18 kDa. Expression profile showed that EJO1 mRNA is highly expressed in the heart, intestine, and ovary of the crab, while there is little or no expression in muscle and hepatopancreas. The differential expression of EJO1 at the different developmental stages of the ovary was further confirmed by Northern blot analysis. In conclusion, EJO1 is a novel gene differentially expressed in the ovary of the Chinese mitten crab, which may play an important role in the ovary development.     

A layover in Europe: Reconstructing the invasion route of asexual lineages of a New Zealand snail to North America

Non‐native invasive species are threatening ecosystems and biodiversity worldwide. High genetic variation is thought to be a critical factor for invasion success. Accordingly, the global invasion of a few clonal lineages of the gastropod Potamopyrgus antipodarum is thus both puzzling and has the potential to help illuminate why some invasions succeed while others fail. Here, we used SNP markers and a geographically broad sampling scheme (N = 1617) including native New Zealand populations and invasive North American and European populations to provide the first widescale population genetic assessment of the relationships between and among native and invasive P. antipodarum. We used a combination of traditional and Bayesian molecular analyses to demonstrate that New Zealand populations harbour very high diversity relative to the invasive populations and are the source of the two main European genetic lineages. One of these two European lineages was in turn the source of at least one of the two main North American genetic clusters of invasive P. antipodarum, located in Lake Ontario. The other widespread North American group had a more complex origin that included the other European lineage and two New Zealand clusters. Altogether, our analyses suggest that just a small handful of clonal lineages of P. antipodarum were responsible for invasion across continents. Our findings provide critical information for prevention of additional invasions and control of existing invasive populations and are of broader relevance towards understanding the establishment and evolution of asexual populations and the forces driving biological invasion.     

Range Expansion in the Northeast Pacific by an Estuary Mud Crab – A Molecular Study

The potential habitat of the introduced estuary crab Rhithropanopeus harrisi (Gould, 1841) in Oregon and central California was surveyed to discover current sites of presence. In order to infer the direction of range expansion and attempt to trace the source population, a 523 bp sequence of the mitochondrial gene Cytochrome c oxidase I was compared between three Pacific populations (28–39 individuals/population), with one mid-Atlantic population included for assessment as a hypothetical source. Notably, this species persists at fairly low densities in the Pacific despite its apparent habitat tolerance, and range expansion and establishment over a broad geographic area. It was possible to deduce that the current Pacific populations descend from a single introduction event. Despite a genetic bottleneck, the historical record of range expansion in a northward direction is reinforced by the molecular data. In order to consider the plausibility of coastal advective transport versus ballast water or aquaculture transport, the new genetic information is evaluated together with the records of northward range expansion for this and other species introduced into San Francisco Bay, including the European green crab, which has had a similar population trajectory. The genetic and temporal patterns that should be expected under these contrasting mechanisms are discussed for typical NE Pacific oceanic conditions, and during years influenced by the El Niño-Southern oscillation.     

Identification, mRNA expression and characterization of a novel ANK-like gene from Chinese mitten crab Eriocheir japonica sinensis

Ankyrins are a family of adapter molecules mediating linkages between integral membrane and cytoskeletal proteins. Ankyrin repeat is one of the most frequently observed amino acid motifs in protein databases. A novel ANK-like gene of Chinese mitten crab Eriocheir japonica sinensis (denoted as EjsANK) was identified and cloned by expressed sequence tag and rapid amplification of cDNA end approaches. The full-length cDNA of EjsANK is 4375 bp and contains an open reading frame of 1095 bp which encodes a 364 amino acids polypeptide (40.23 kD) bearing seven ankyrin repeats. EjsANK cDNA has a 3073 bp uniquely long 3′ untranslated region with three K-box elements, one GY-like box domain and one Brd-like box domain. Sequence alignment and three-dimensional structural analyses revealed that EjsANK should be a novel cytosolic member of the ankyrin family. Fluorescent real-time quantitative RT-PCR approach was performed to examine the expression profiles of EjsANK mRNA by testing its relative level in three types of tissues at three different developmental stages, respectively. We found that the relative level of EjsANK mRNA expression was significantly higher in the abdomen at the first crab stage. Functional bioinformatics prediction analyses indicated that EjsANK has an analogical effect like IκB which is a key component of IκB/NF-κB complex in mammalian cells playing very important roles in the development process. Results suggest that EjsANK gene is involved in the early developmental regulation of Chinese mitten crab, especially brachyurization regulation.     

Patterns of nuclear and mitochondrial DNA variation in Iberian populations of <Emphasis Type="Italic">Emys orbicularis</Emphasis> (Emydidae): conservation implications

The European pond turtle (Emys orbicularis) is threatened and in decline in several regions of its natural range, due to habitat loss combined with population fragmentation. In this work, we have focused our efforts on studying the genetic diversity and structure of Iberian populations with a fine-scale sampling (254 turtles in 10 populations) and a representation from North Africa and Balearic island populations. Using both nuclear and mitochondrial markers (seven microsatellites, ∼1048 bp nDNA and ∼1500 bp mtDNA) we have carried out phylogenetic and demographic analyses. Our results show low values of genetic diversity at the mitochondrial level although our microsatellite dataset revealed relatively high levels of genetic variability with a latitudinal genetic trend decreasing from southern to northern populations. A moderate degree of genetic differentiation was estimated for Iberian populations (genetic distances, F _ST values and clusters in the Bayesian analysis). The results in this study combining mtDNA and nDNA, provide the most comprehensive population genetic data for E. orbicularis in the Iberian Peninsula. Our results suggest that Iberian populations within the Iberian–Moroccan lineage should be considered as a single subspecies with five management units, and emphasize the importance of habitat management rather than population reinforcement (i.e. captive breeding and reintroduction) in this long-lived species.     

Microsatellite variation in Bavarian populations of European grayling (Thymallus thymallus): Implications for conservation

European grayling populations in Bavaria have shown steady declines during the last 10–20years. In order to provide guidelines for conservation strategies and future management programs, we investigated the genetic structure of 15 grayling populations originating from three major Central European drainages (the Danube, the Elbe and the Rhine/Main) using 20 microsatellite loci. Genetic divergence between the three drainage systems was substantial as illustrated by highly significant heterogeneity of genotype frequencies, high number of drainage-specific private alleles, high between-drainage F _ST values, high assignment success of individuals to their drainage of origin and the high bootstrap support for the genetic distance based drainage-specific population clusters. In agreement with earlier studies, microsatellites revealed relatively low levels of intrapopulational genetic diversity in comparison to the overall level of variation across populations. Maximum likelihood methods using the coalescent approach revealed that the proportion of common ancestors was generally high in native populations and that the estimates of N _e were correlated with the genetic diversity parameters in all drainages. The number of effective immigrants per generation (N _e m) was less than one for all pairwise comparisons of populations within the drainages, indicating restricted interpopulational gene flow. Based on these findings we recommend a drainage and sub-drainage specific conservation of grayling populations in order to preserve their overall genetic diversity and integrity. For large-scale stocking actions to supplement declining or to restore extinct populations, creation of separate broodstocks for major conservation units (ESUs and MUs) is warranted. This revised version was published online in July 2006 with corrections to the Cover Date.     

Introduced plants of the invasive Solidago gigantea (Asteraceae) are larger and grow denser than conspecifics in the native range

Introduced plant species that became successful invaders appear often more vigorous and taller than their conspecifics in the native range. Reasons postulated to explain this better performance in the introduced range include more favourable environmental conditions and release from natural enemies and pathogens. According to the Evolution of Increased Competitive Ability hypothesis (EICA hypothesis) there is a trade‐off between investment into defence against herbivores and pathogens, and investment into a stronger competitive ability. In this study, we conducted field surveys to investigate whether populations of the invasive perennial Solidago gigantea Ait (Asteraceae) differ with respect to growth and size in the native and introduced range, respectively. We assessed size and morphological variation of 46 populations in the native North American range and 45 populations in the introduced European range. Despite considerable variation between populations within continents, there were pronounced differences between continents. The average population size, density and total plant biomass were larger in European than in American populations. Climatic differences and latitude explained only a small proportion of the total variation between the two continents. The results show that introduced plants can be very distinct in their growth form and size from conspecifics in the native range. The apparently better performance of this invasive species in Europe may be the result of changed selection pressures, as implied by the EICA hypothesis.     

Invasion history of North American Canada thistle,Cirsium arvense

Aim Canada thistle (Cirsium arvense– Cardueae, Asteraceae) is one of the worst invasive plants world‐wide. Native to Eurasia, its unintentional introduction into North America now threatens the native flora and is responsible for enormous agricultural losses. The goals of this study are to: (1) reconstruct the evolutionary history of C. arvense and estimate how often it may have colonized North America, (2) compare the genetic diversity between European and North American populations to detect signs of demographic bottlenecks and/or patterns of population admixture, and (3) conduct bioclimatic comparisons to infer eventual niche shifts following this species’ introduction into North America. Location Europe and North America. Methods A total of 1522 individuals from 58 populations were investigated with six microsatellite markers. Estimates of heterozygosity (H_E) and allelic richness (R_S) were quantified for each population, and population structure was inferred via analyses of molecular variance (AMOVAs), principal components analyses (PCAs), Mantel tests and Bayesian clustering analyses. Climatic niche spaces were based on 19 bioclimatic variables extracted from approximately 32,000 locations covering the entire range, and compared using PCA and hierarchical cluster analysis. Results Although there is evidence of multiple introductions from divergent European lineages, North American populations of C. arvense exhibited significantly lower levels of genetic diversity than their putative ancestors. Bioclimatic comparisons pointed to a high degree of niche conservatism during invasion, but indicated that genotypes from the former USSR and Central European mountain chains were probably best adapted to invade North America upon entry into the continent. Main conclusions Genetic and historical data suggest that C. arvense first entered North America from Western Europe with the first European settlers, and was later introduced from Eastern Europe into the prairie states during the agricultural boom. The species went through a significant bottleneck following its introduction into the New World, but the level of genetic diversity remained high owing to admixture between genetically differentiated lineages and to a highly efficient outcrossing breeding system.     

Contrasting mitochondrial diversity of European starlings (Sturnus vulgaris) across three invasive continental distributions

European starlings (Sturnus vulgaris) represent one of the most widespread and problematic avian invasive species in the world. Understanding their unique population history and current population dynamics can contribute to conservation efforts and clarify evolutionary processes over short timescales. European starlings were introduced to Central Park, New York in 1890, and from a founding group of about 100 birds, they have expanded across North America with a current population of approximately 200 million. There were also multiple introductions in Australia in the mid‐19th century and at least one introduction in South Africa in the late 19th century. Independent introductions on these three continents provide a robust system to investigate invasion genetics. In this study, we compare mitochondrial diversity in European starlings from North America, Australia, and South Africa, and a portion of the native range in the United Kingdom. Of the three invasive ranges, the North American population shows the highest haplotype diversity and evidence of both sudden demographic and spatial expansion. Comparatively, the Australian population shows the lowest haplotype diversity, but also shows evidence for sudden demographic and spatial expansion. South Africa is intermediate to the other invasive populations in genetic diversity but does not show evidence of demographic expansion. In previous studies, population genetic structure was found in Australia, but not in South Africa. Here we find no evidence of population structure in North America. Although all invasive populations share haplotypes with the native range, only one haplotype is shared between invasive populations. This suggests these three invasive populations represent independent subsamples of the native range. The structure of the haplotype network implies that the native‐range sampling does not comprehensively characterize the genetic diversity there. This study represents the most geographically widespread analysis of European starling population genetics to date.     

Scaling the consequences of interactions between invaders from the individual to the population level

The impact of human‐induced stressors, such as invasive species, is often measured at the organismal level, but is much less commonly scaled up to the population level. Interactions with invasive species represent an increasingly common source of stressor in many habitats. However, due to the increasing abundance of invasive species around the globe, invasive species now commonly cause stresses not only for native species in invaded areas, but also for other invasive species. I examine the European green crab Carcinus maenas, an invasive species along the northeast coast of North America, which is known to be negatively impacted in this invaded region by interactions with the invasive Asian shore crab Hemigrapsus sanguineus. Asian shore crabs are known to negatively impact green crabs via two mechanisms: by directly preying on green crab juveniles and by indirectly reducing green crab fecundity via interference (and potentially exploitative) competition that alters green crab diets. I used life‐table analyses to scale these two mechanistic stressors up to the population level in order to examine their relative impacts on green crab populations. I demonstrate that lost fecundity has larger impacts on per capita population growth rates, but that both predation and lost fecundity are capable of reducing population growth sufficiently to produce the declines in green crab populations that have been observed in areas where these two species overlap. By scaling up the impacts of one invader on a second invader, I have demonstrated that multiple documented interactions between these species are capable of having population‐level impacts and that both may be contributing to the decline of European green crabs in their invaded range on the east coast of North America.     

Evolution of an invasive phenotype: shift to belowground dominance and enhanced competitive ability in the introduced range

In response to novel selection pressures in an introduced range, non-native species may evolve more competitive phenotypes unique from those of their native range. We examined the existence of an invasive phenotype in the herbaceous perennial Artemisia vulgaris, a frequent invader of the Northeast and Mid-Atlantic US. Populations from both the native (European) and the introduced (North American) ranges were grown in intra-specific competition (same population), inter-specific competition with the native perennial herb Solidago canadensis, and alone in a common garden to quantify shifts in resource allocation and neighbor effects on performance and competitive ability. Without competition, introduced A. vulgaris populations were much shorter than native populations, but germinated earlier, produced more ramets, more belowground and total biomass, and maintained higher root-to-shoot ratios. Under inter- and intra-specific competitions, introduced A. vulgaris populations were shorter, but produced more ramets, belowground, and total biomass than native populations. S. canadensis belowground and total biomass were more highly suppressed by introduced than native A. vulgaris. Our data suggest that since the introduction to North America, A. vulgaris has evolved a more competitive invasive phenotype characterized by many short ramets with more extensive root/rhizome networks. This rapid evolutionary shift likely benefits A. vulgaris in its introduced range by allowing establishment and subsequent dominance in dense stands of existing vegetation.     

Secondary contact and local adaptation contribute to genome‐wide patterns of clinal variation in Drosophila melanogaster

Populations arrayed along broad latitudinal gradients often show patterns of clinal variation in phenotype and genotype. Such population differentiation can be generated and maintained by both historical demographic events and local adaptation. These evolutionary forces are not mutually exclusive and can in some cases produce nearly identical patterns of genetic differentiation among populations. Here, we investigate the evolutionary forces that generated and maintain clinal variation genome‐wide among populations of Drosophila melanogaster sampled in North America and Australia. We contrast patterns of clinal variation in these continents with patterns of differentiation among ancestral European and African populations. Using established and novel methods we derive here, we show that recently derived North America and Australia populations were likely founded by both European and African lineages and that this hybridization event likely contributed to genome‐wide patterns of parallel clinal variation between continents. The pervasive effects of admixture mean that differentiation at only several hundred loci can be attributed to the operation of spatially varying selection using an F_ST outlier approach. Our results provide novel insight into the well‐studied system of clinal differentiation in D. melanogaster and provide a context for future studies seeking to identify loci contributing to local adaptation in a wide variety of organisms, including other invasive species as well as temperate endemics.     

Development and characterization of new microsatellite loci from Chinese mitten crab (Eriocheir sinensis)

A set of new microsatellite loci were isolated from Chinese mitten crab and characterized in two Chinese mitten crab populations. The number of the alleles ranged from 3 to 8 and observed and expected heterozygosity varied from 0.0870 to 0.9565 and 0.3092 to 0.8367 in each population, respectively. Loci HLJEsin1, HLJEsin4, HLJEsin30 in CLN population and HLJEsin6 in CJS population presented significant deviation from Hardy–Weinberg equilibrium (PHWE < 0.001). These markers will be useful for population studies.