Isolation and characterization of microsatellite markers in the newly discovered invasive fruit fly pest in Africa, Bactrocera invadens (Diptera: Tephritidae)

We describe the isolation and characterization of 11 polymorphic microsatellite loci from the recently discovered fruit fly pest, Bactrocera invadens. The polymorphism of these loci was tested in individual flies from two natural populations (Sri Lanka and Democratic Republic of Congo). Allele number per locus ranged from three to 15 and eight loci displayed a polymorphic information content greater than 0.5. These microsatellite loci provide useful markers for studies of population dynamics and invasion history of this pest species.     

Polymorphic microsatellite DNA markers from the Oriental Fruit Fly Bactrocera dorsalis (Hendel)

Six polymorphic microsatellite loci are isolated from the Oriental fruit fly Bactrocera dorsalis (Hendel), an agricultural pest in Asia, including Taiwan. To assess their potential utility as high‐resolution genetic markers, polymerase chain reaction (PCR) primers, amplification conditions, and an automated fluorescence detection protocol were developed. In analyses of 71 individual flies from six different areas of Taiwan, allele numbers ranged from five to 25 were detected for each locus. The observed heterozygosity ranged between 0.268 and 0.737 among these loci. No linkage disequilibrium was found. These microsatellite markers have potential utility to population structure and gene flow studies of B. dorsalis (Hendel).     

Microsatellite analysis revealed a different approach of control of olive fly population (Bactrocera oleae) in Slovenia

The olive fruit fly (Bactrocera oleae Gmelin) is the most important olive pest in the north‐eastern Adriatic coast region. Despite the importance of olive production in the region, and the significance of the olive fruit fly, no information with respect to genetic diversity, population structure or dispersion patterns of this pest is available. The aims of this study were to investigate the genetic structure of the olive fly population in the Slovenian Istria region using microsatellite markers to determine olive fruit fly migration between locations and to establish an appropriate and effective strategy for controlling the pest population. Analysis was performed on a sample of 117 flies, collected from attacked olive fruits at three different locations. Olive fruit flies were genotyped using eight microsatellite loci. Sixty‐six alleles were identified over all microsatellite loci with an average of 8.25 alleles per locus. The population structure was determined with methods based on Bayesian principles using the BAPS 6.0 and STRUCTURE 2.3 programs. Genetic analysis confirmed unlimited migration and random mating between individuals of different microlocations, which suggests time‐coordinated first treatment in the region would be the best solution.     

New Polymorphic Microsatellites in Glossina pallidipes (Diptera: Glossinidae) and Their Cross-Amplification in Other Tsetse Fly Taxa

We report the development and characterization of three new microsatellite markers in the tsetse fly, Glossina pallidipes (Diptera: Glossinidae). Fifty-eight alleles were scored in 192 individuals representing six natural populations. Allelic diversity ranged from 9 to 28 alleles per locus (mean 19.3 +/- 5.5). Averaged across loci, observed heterozygosity was 0.581 +/- 0.209, and expected heterozygosity was 0.619 +/- 0.181. Cross-species amplifications of the G. pallidipes loci in other tsetse fly taxa are reported.     

Characterization of microsatellite loci in the house fly,Musca domestica L. (Diptera: Muscidae)

The house fly, Musca domestica L., is a cosmopolitan species with a capacity to transmit human pathogens. Here, we report on the development of polymorphic microsatellite loci for house flies and present preliminary results from four house fly subpopulations from Manhattan, Kansas. Twenty‐four microsatellite loci were isolated and characterized using DNA enriched for repeat sequences. Forty individuals from four locations in Kansas were assayed to identify for polymorphic loci. Eight loci were polymorphic with the number of alleles ranging from three to six.     

Polymorphic microsatellite DNA markers in Bactrocera cacuminata (Hering) (Diptera: Tephritidae)

Bactrocera cacuminata (Hering) is a nonpest member of the Bactrocera dorsalis complex offering a platform to check hypotheses extrapolated from the more studied pest species. Six polymorphic microsatellite loci were isolated from enriched genomic libraries constructed using a biotin/streptavidin capture protocol. Allele number varied between three and nine; the expected heterozygosity ranged between 0.29 and 0.81. No significant deviations from Hardy–Weinberg equilibrium or linkage disequilibrium were found. These microsatellite markers have potential to be used to examine population structure and mating systems in this tropical fruit fly.     

Parasites that attack stable fly and house fly (Diptera: Muscidae) puparia during the winter on dairies in northwestern Florida

Throughout the winter and early spring months, stable fly, Stomoxys calcitrans (L.), and house fly, Musca domestica L., puparia were collected from silage, hay, and manure from six dairies in northwestern Florida and evaluated for parasitism. Of the puparia producing flies or parasites, 23% of the stable flies and 46% of the house flies were parasitized. The predominant parasite observed attacking muscoid flies (76% for stable flies and 58% for house flies) was Spalangia cameroni Perkins. Muscidifurax sp. was recovered from 11 and 36% of the stable fly and house fly pupae, respectively. Other parasite species encountered were Spalangia endius Walker and S. nigroaenea Curtis. Significantly more parasitized fly pupae were collected from silage than from hay residues or manure. Winter and early spring parasite populations in northwestern Florida appear to be present as long as viable fly pupae are available to support the developing parasites.     

Winter feeding sites of hay in round bales as major developmental sites of Stomoxys calcitrans (Diptera: Muscidae) in pastures in spring and summer

The stable fly, Stomoxys calcitrans (L.), historically has been a pest of livestock in confined operations but seldom of animals on pastures or rangelands. In the past two decades, however, S. calcitrans has become a major pest of cattle and horses on pastures in the midwestern United States. Although there usually is an overabundance of diverse stable fly and house fly, Musca domestica L., larval habitats in confined livestock operations, no larval habitat for stable flies has been clearly identified in the pasture-range environment. Because the winter feeding of hay in round bales results in significant amounts of hay wastage that when mixed with manure, might develop into suitable larval habitats, this study evaluated these areas as developmental sites for the abundant stable flies in pastures. There was a trend for fly traps placed in the vicinity of hay feeding sites to catch more stable flies than those placed distant from these sites. Estimates of stable flies emerging from these sites ranged from 102 to 1225 flies per core sample (25 by 25 cm). The mean number of adult stable flies during May and June 2001 through 2004 correlated negatively with the average minimum temperatures during the preceding winter (November-February) but not with rainfall or temperatures during the spring. These results support the hypothesis that winter feeding sites of hay in round bales are the main source of stable flies in pastures.     

Trinucleotide microsatellite loci in the yellow dung fly Scathophaga stercoraria (Diptera: Scathophagidae)

We describe six polymorphic, trinucleotide microsatellite loci that were isolated from the yellow dung fly Scathophaga stercoraria (Diptera: Scathophagidae), a model system for examining the mechanisms behind sperm competition in species with internal fertilization. These microsatellites yielded between 3 and 11 alleles per locus in a sample of 20–48 dung flies collected from several sites in Cheshire, UK. Observed and expected heterozygosities varied between 0.195 and 0.900 and 0.335–0.842, respectively. These markers should allow us to investigate postcopulatory sexual selection processes in this fly much more fully than has been possible in the past.     

Relative attractiveness of paired BL and BLB fluorescent bulbs for house and stable flies (Diptera: Muscidae)

Blacklight (BL) and blacklight blue (BLB) fluorescent bulbs were combined in an electrocuting fly trap and compared with BL or BLB bulbs alone for fly attraction. Combinations of BL and BLB bulbs did not attract more house flies, Musca domestica, or stable flies, Stomoxys calcitrans than were attracted by BL bulbs used alone.     

PERMANENT GENETIC RESOURCES: Polymorphic microsatellite loci for Diachasma alloeum (Hymenoptera: Braconidae)

Here, we report the isolation of 21 novel primers for amplification of microsatellite loci in Diachasma alloeum (Hymenoptera: Braconidae). Diachasma alloeum is a larval parasitoid of the apple maggot fly (Rhagoletis pomonella), which is an economically significant agricultural pest species and a textbook example of sympatric speciation via host-plant shifting. These microsatellite markers will prove useful both for assessing genetic relationships between different host-associated populations of D. alloeum, as well as for future R. pomonella biological control programmes. We also report the cross-species amplification of several loci for Diachasmimorpha mellea and Diachasma ferrugineum, parasitoids of R. pomonella and R. cingulata, respectively.     

Polymorphic microsatellite markers for population analysis of a tephritid pest species, Bactrocera tryoni

To obtain a set of microsatellite markers for the Queensland fruit fly Bactrocera tryoni , a genomic library was screened with a number of simple repeat oligonucleotide probes. Sequencing recovered 22 repeat loci. The microsatellite sequences were short, with repeat numbers ranging from five to 11. Of these, 16 polymerase chain reaction (PCR) primer sets yielded amplifiable products, which were tested on 53 flies from five widely separated sites. All loci showed polymorphism in the population sample, with the number of alleles ranging from two to 16. Several dinucleotide repeats showed alleles separated by single-base differences and multiple steps, suggesting a mutation process more complex than the stepwise mutation model.     

Effects of stable flies (Diptera: Muscidae) on weight gains of grazing yearling cattle.

Differences in weight gains caused by stable flies, Stomoxys calcitrans (L.), on grazing yearling steer/calves averaged 0.2 kg per steer in a 3-yr study on canyon range pastures in West Central Nebraska, Stable fly numbers averaged 0.85 per front leg on treated calves and 3.64 per front leg on control calves. In 2 of the 3 yr after the grazing trials were completed, the calves were placed in a feedlot and fed a finishing ration. Compensatory gain did not occur in the feedlot after the stable fly stress was removed.     

Larvicidal activity of endectocides against pest flies in the dung of treated cattle

Cattle were treated with topical formulations of endectocides to assess the larvicidal activity of faecal residues against horn fly, Haematobia irritans (L.), house fly, Musca domestica L., and stable fly, Stomoxys calcitrans (L.) (Diptera: Muscidae). In laboratory bioassays, doramectin, eprinomectin and ivermectin suppressed horn fly in dung of cattle treated at least 4 weeks previously and suppressed house fly and stable fly in dung of cattle treated 1-5 weeks previously. Moxidectin suppressed horn fly in dung from cattle treated no more than one week previously and did not suppress house fly and stable fly. Results combined for the three species across two experiments suggested that, ranked in descending order of larvicidal activity, doramectin > ivermectin approximately = eprinomectin > moxidectin.     

Isolation and Characterization of Microsatellite Loci for Striped Bass (Morone saxatilis)

Genetic variation has been difficult to detect in striped bass (Morone saxatilis). Therefore, we identified and characterized 13 microsatellite loci to provide additional genetic markers for striped bass. Microsatellites were identified by screening a striped bass genomic library or by using primers developed for European sea bass (Dicentrarchus labrax) microsatellite loci. We found that 6 of the 13 microsatellite loci were polymorphic in DNA samples obtained from wild populations of striped bass. The number of alleles per locus varied from 3 to 12, and the observed heterozygosities ranged from 0.55 to 0.78. These results indicate that microsatellite loci provide more alleles and higher heterozygosities than other genetic markers developed for striped bass. Received November 9, 1999; accepted February 11, 2000.     

Isolation and characterization of 11 microsatellite loci from the black fly, Simulium negativum (Diptera: Simuliidae)

Eleven polymorphic microsatellite loci were isolated and developed from the black fly, Simulium negativum, a member of the Simulium arcticum sibling species complex. The observed heterozygosity of the 11 loci ranged from 0.03 to 0.83. The number of alleles per locus ranged from 8 to 19. Significant linkage disequilibria were encountered only for the primer pairs BF7-1 with BF7-5 and BF6-32 with BF7-16. Presumably, these microsatellite loci can be used to study genetic structure within the entire S. arcticum complex.     

New diets for production of house flies and stable flies (Diptera: Muscidae) in the laboratory.

A diet for rearing the house fly, Musca domestica (L.), was developed from feed constituents available on a year-round basis in Gainesville, FL. The diet, called the Gainesville House Fly Diet, performed as well or better than the Chemical Specialties Manufacturers' Association fly larval medium (CSMA) and can be mixed, bagged, and delivered by a local feed mill within 3 d. By adding pelleted peanut hulls 1:1 by volume, the house fly diet becomes suitable for rearing the stable fly, Stomoxys calcitrans (L.). Use of these diets and the economics involved are discussed further.     

Isolation, characterization, inheritance and linkage of microsatellite markers in Tetranychus kanzawai (Acari: Tetranychidae)

We isolated and characterized seven microsatellite markers in Tetranychus kanzawai (Acari: Tetranychidae). We also examined the conformity of the isolated markers to Mendelian laws and analyzed linkage among the microsatellite loci. All microsatellite markers fit expected 1:1 disomic segregation ratio and hence were inherited in a Mendelian manner. Significant pairwise linkage was detected in three pairs of microsatellite loci. These isolated microsatellite markers may become a powerful tool for the study of behavioral ecology, population genetics, and genome mapping of T. kanzawai.     

Seasonal abundance of stable flies and filth fly pupal parasitoids (Hymenoptera: Pteromalidae) at Florida equine facilities

Beginning in November 2007 and continuing until December 2009, weekly stable fly, Stomoxys calcitrans (L.), surveillance was conducted at four equine facilities near Ocala, FL, by using alsynite sticky traps for adults and by searching immature developmental sites for pupae. Adult stable fly trap captures were highly variable throughout the year, ranging from 0 to 1,400 flies per trap per farm. The greatest adult stable fly activity was observed during the spring months of March and April, with weekly three-trap means of 121 and 136 flies per farm, respectively. The importance of cultural control measures was most apparent on the only farm with no reported insecticide use and the lowest stable fly trap captures, where an intense daily sanitation and composting program was conducted. A survey of on-site filth fly pupae revealed that 99.9% of all parasitoids recovered were Spalangia spp., consisting of Spalangia cameroni Perkins (56.5%), Spalangia nigroaenea Curtis (34.0%), Spalangia endius Walker (5.8%), and Spalangia nigra Latreille (3.7%). The implications of these findings are discussed.     

Isolation and Characterization of Twenty Microsatellite Loci in Japanese Sea Cucumber (Stichopus japonicus)

Twenty microsatellite markers were first developed from the Japanese sea cucumber Stichopus japonicus using an enrichment protocol. Of the 20 microsatellite loci, 19 loci were polymorphic in the population examined. At these polymorphic loci, the number of alleles per locus varied from 2 to 15, and the observed heterozygosities ranged from 0.03 to 0.97, which is considerably higher than those previously found for allozymes. The high variability of the microsatellite markers identified in this study will make them excellent tools for genetic analyses of S. japonicus.