Characterization of polymorphic microsatellite markers for the blowfly Chrysomya albiceps (Diptera: Calliphoridae)

Chrysomya albiceps is a blowfly of great medical, sanitary and forensic importance widely distributed in the Afrotropical, southern Palaearctic, northern Oriental regions and, recently, in Central and South Americas. Here, we report the characterization of 13 polymorphic microsatellite markers for C. albiceps. The number of alleles ranged from three to 13 alleles with expected heterozygosities ranging from 0.4668 to 0.8408. These markers will be extremely useful for investigating many important aspects of this species such as population structure, dispersal and colonization dynamics.     

Development of polymorphic microsatellite markers for the sorghum plant bug, Stenotus rubrovittatus (Heteroptera: Miridae)

Nine polymorphic microsatellite DNA markers from the sorghum plant bug, Stenotus rubrovittatus, were isolated and characterized. These markers were used to analyse 22 individuals from a single field population. The number of alleles at these nine loci ranged from two to 28 (mean = 11.4) and heterozygosity ranged from 0.27 to 0.86 (mean = 0.58). Stenotus rubrovittatus has shown rapid population growth in the decades since the first report in the 1980s of serious damage to a rice crop. These microsatellite markers will be of value for studying both the population genetics and population dynamics of S. rubrovittatus.     

Experimental skin lesions from larvae of the bot fly Dermatobia hominis

Skin biopsies from larvae of Rattus norvegicus, experimentally infested with Dermatobia hominis (Linnaeus Jr) (Diptera: Cuterebridae), were processed for histopathological studies. Two days after infestation, the first-stage larvae (L1) were located deep in the dermis, surrounded by an inflamed area infiltrated predominantly by neutrophils. On the fourth day a thin necrotic layer could be seen close to the larvae, surrounded by large numbers of neutrophils, lymphocytes, macrophages with a few eosinophils and mast cells. A small warble was formed after the fourth day, increasing in size until the seventh day, when the L1 moulted to the second-stage larva (L2). The inflammatory process continued with increasing numbers of neutrophils, macrophages, lymphocytes, eosinophils and mast cells invading the area, as well as the proliferation of fibroblasts and endothelial cells and the appearance of a few localized haemorrhages. After 18-20 days, the L2 moulted to the third-stage larva (L3), when a few plasma cells could be seen in the inflamed area. At 25-30 days there was a reduction in the necrotic layer, as well as in the number of neutrophils and lymphocytes, although large amounts of eosinophils, plasma cells, and collagen fibres were seen. The L3 usually left the host after 30 days. Two days later, the larval cavity was reduced, mast cells infiltrated the region and collagen fibre production were increased. After 7 days, an intense infiltration of plasma cells and scattered necrotic areas could be seen. A scar formed after 10 days. This study showed the laboratory rat to be a suitable model for studies of D. hominis infestation.     

Twenty-three new microsatellite loci in the stable fly, Stomoxys calcitrans (L.) (Diptera: Muscidae)

The stable fly, Stomoxys calcitrans (L.), is a significant pest of cattle. Twenty-three microsatellite markers were isolated from a repeat-enriched genomic library of S. calcitrans. We characterized variation at these markers and found that 17 loci were polymorphic in two fly populations from Florida. Two to nine alleles were observed among the variable microsatellite loci and expected heterozygosities ranged from 0.03704 to 0.85115. These markers will be useful for characterizing population genetic differentiation and for tracking the migration patterns of stable flies in the USA and worldwide.     

Isolation of polymorphic microsatellite markers in the tree sparrow (Passer montanus)

To enable the study of the population genetics of the tree sparrow (Passer montanus), which is distributed in many islands of Japan, nine polymorphic microsatellite loci were isolated from 32 individuals. The number of alleles per locus ranged from 2 to 17, and observed heterozygosities ranged from 0.1250 to 0.9375. In eight of the nine loci, the heterozygosities were not significantly different from those expected from Hardy-Weinberg equilibrium. Cross-species amplification in the russet sparrow (P. rutilans) was successful with all primer sets, which were highly polymorphic, suggesting these markers are useful for the population genetics of genus Passer.     

Characterization of polymorphic microsatellite DNA markers in the black-tailed godwit (Limosa limosa: Aves)

We isolated and tested 16 microsatellite loci in black-tailed godwits from the Netherlands (Limosa limosa limosa), and from Australasia (subspecies melanuroides). One locus was monomorphic, two loci had null-alleles and one was significantly heterozygote deficient. The remaining 12 polymorphic loci had on average 7.9 alleles (range 5-11) and the mean expected heterozygosity was 0.69. No significant linkage disequilibrium between the loci was observed and all loci were autosomal. Fourteen loci were successfully cross-amplified in bar-tailed godwit (Limosa lapponica).     

Interspecific microsatellite markers for the study of pinniped populations

Microsatellites have rapidly become the marker of choice for a wide variety of population genetic studies. Here we describe 20 pinniped microsatellite markers which have been tested across 18 pinniped species. The majority of these markers have broad utility in all pinnipeds and provide a strong base for detailed population genetic studies in the Pinnipedia.     

Isolation and multiplex genotyping of polymorphic microsatellite DNA markers in the snakehead murrel, Channa striata

Seven polymorphic microsatellite loci were isolated and characterized for the snakehead murrel, Channa striata (Channidae), a valuable tropical freshwater fish species. Among 25 specimens collected from Kedah state in Malaysia, the number of alleles per locus ranged from 2 to 7. Observed and expected heterozygosities ranged from 0.120 to 0.880 and 0.117 to 0.698, respectively. A single locus (CS1-C07) was significantly deviated from Hardy-Weinberg equilibrium after Bonferroni correction. These novel markers would be useful for population genetic studies of the C. striata.     

Nine polymorphic microsatellite loci for the parasitic wasp Neotypus melanocephalus (Hymenoptera: Ichneumonidae)

Nine polymorphic microsatellite loci were isolated from Neotypus melanocephalus (Gmelin), a parasitoid of the parasitic large blue butterfly Maculinea nausithous. Allelic diversity and heterozygosity were quantified in samples from the Upper Rhine valley in Southwest Germany.     

Isolation and characterization of 19 polymorphic microsatellite DNA markers in the Japanese brown frog (Rana japonica)

We report the isolation and characterization of 19 polymorphic microsatellite DNA markers in the Japanese brown frog (Rana japonica). These markers were tested in 24 individuals each collected from three distinct populations in Ichikai-machi, Tochigi Prefecture. The number of observed alleles per locus ranged from 3 to 24 across all populations, and the values of observed and expected heterozygosities ranged from 0.130 to 1 and from 0.125 to 0.941, respectively. These microsatellite loci will be useful for investigating the intraspecific genetic variation and population structure of this species.     

Development of polymorphic microsatellite markers in Hessian fly, Mayetiola destructor (Say)

A microsatellite library was prepared from size-selected genomic DNA of Hessian fly (Mayetiola destructor). Approximately 81% of recovered clones hybridized with microsatellite motif-specific probes. Subsequently, 2350 clones were sequenced. Sixty-two individual flies from laboratory strains were used to test for reliability and polymorphism in 50 of the microsatellites by gel electrophoresis; 18 were further tested with capillary electrophoresis. Of these, 17 behaved as a polymorphic single locus appropriate for population analysis.     

Isolation and characterization of polymorphic microsatellite DNA markers in the sea squirt Halocynthia roretzi

The sea squirt Halocynthia roretzi is an important marine food resource species that is found in the waters around Korea. We describe the isolation and characterization of 13 new polymorphic microsatellite loci in 96 sea squirt samples that were collected from the marine environment of Samcheok on the east coast of Korea. The number of alleles that were observed for each locus ranged from six to 32, and the value of expected and observed heterozygosities was 0.504-0.922 and 0.396-0.813, respectively. These markers will be useful tools for future population studies.     

Isolation and characterization of polymorphic microsatellite markers from Asian malaria mosquito Anopheles sinensis (Diptera: Culicidae)

Microsatellite-containing region were isolated and characterized in Anopheles sinensis, a primary vector of malaria parasites in Asia. An enrichment protocol yielded 252 microsatellite sequences. We designed primers to amplify 20 unique microsatellites, 14 of which amplify cleanly and were polymorphic. A survey of 24 individuals showed that 12 loci are highly variable with the number of alleles ranging from two to 11, and expected heterozygosity ranging from 0.116 to 0.903. These markers will be useful for population genetic studies and genome mapping in A. sinensis.     

Development and characterization of 16 polymorphic microsatellite loci in the Tibet endemic fish Glyptosternum maculatum

Sixteen novel polymorphic microsatellite loci were developed and characterized in Glyptosternum maculatum, an endemic fish inhabiting the Yarlung Zangbo River valley, Tibet, China. These loci were developed after the enrichment of microsatellite-containing genomic DNA fragments with a subtractive hybridization protocol. The polymorphism of these loci was analysed with 36 individuals representing a geographical population. The number of alleles found at these loci ranged from 3 to 10. The observed heterozygosity and expected heterozygosity ranged from 0 to 1 and from 0.31 to 0.75, respectively. These microsatellite loci will certainly facilitate the determination of the genetic structure of G. maculatum and the conservation of its genetic resource.     

Development and characterization of eight polymorphic microsatellite markers for Daphnia atkinsoni (Crustacea: Ctenodaphnia)

A microsatellite‐enriched genomic library was developed for the water flea Daphnia atkinsoni Baird, 1859, a dominant species of intermittent wetlands in Europe. Eight polymorphic microsatellite markers were successfully optimized. Characterization of 77 individuals from Belgium and Spain showed moderate (in the former) to high (in the latter) levels of polymorphism with two to 11 alleles per locus and an observed heterozygosity ranging from 0 to 0.87. Some of these microsatellite markers were successfully amplified in three other Daphnia species (D. magna n = 4, D. similis n = 6; D. obtusa n = 6).     

Characterization of polymorphic microsatellite markers for a coniferous shrub Juniperus sabina (Cupressaceae)

Juniperus sabina L. is an evergreen coniferous shrub, with a widespread distribution in Asia and Europe. It is one of the key species for restoration of the ecosystem in desertification areas. In the present study, we isolated and characterized eight microsatellite loci of this species. The number of alleles per locus ranged from two to 27, with observed heterozygosity values ranging from 0.256 to 0.744 and expected heterozygosity from 0.276 to 0.939. The markers developed in this study could be useful for population genetics studies of J. sabina.     

Characterization of polymorphic microsatellite markers in the brine shrimp Artemia (Branchiopoda, Anostraca)

The brine shrimp Artemia is a complex genus containing sexual species and parthenogenetic lineages. Artemia franciscana is native to America and its cysts (diapausing eggs) are used worldwide as a food source in aquaculture. As a consequence, this anostracan has become an invasive species in many hypersaline aquatic ecosystems of other continents. Parthenogenetic Artemia lineages occur only in the Old World. Ten and five microsatellite markers were developed to characterize two populations for A. franciscana and two populations for diploid parthenogenetic Artemia, respectively. For A. franciscana the number of alleles ranged from 11 to 58 per locus, while for parthenogens the number of alleles ranged from three to 10. The levels of heterozygosity in A. franciscana and in parthenogens ranged from 0.115 to 0.976 and from 0.000 to 0.971, respectively. These microsatellite loci showed a high population assignment power, which will be useful for future studies of population genetics and invasive processes in Artemia.     

Development of microsatellite markers for the grapevine fungal pathogen Phaeomoniella chlamydospora

Twenty polymorphic microsatellite markers from microsatellite-enriched genomic DNA of the grapevine fungal pathogen, Phaeomoniella chlamydospora, were developed and characterized. The markers were used to genotype isolates from Australia and from Europe/Eurasia. The number of alleles per locus ranged from two to 11. Gene diversity per locus ranged from 0.08 to 0.63 among Australian isolates, and from 0.2 to 0.77 among isolates from Europe/Eurasia, demonstrating the suitability of these markers for population genetic studies of P. chlamydospora. Eighteen of the 20 markers also amplified a product in the closely related Phaeoacremonium aleophilum.     

New polymorphic microsatellite loci for population studies in the European anchovy, Engraulis encrasicolus (L.)

Eleven microsatellite loci were developed in the European anchovy, Engraulis encrasicolus, and tested in samples from two geographically distant populations (Atlantic and Mediterranean Sea). Number of alleles ranged from eight to 28 and observed heterozygosity from 0.440 to 0.920. There was no evidence of linkage disequilibrium, although two loci are indeed linked. All loci were in Hardy-Weinberg equilibrium, except for one locus in the Atlantic and two loci in the Mediterranean sample. These three loci plus two more showed evidence for null alleles.