Christopher J. Geden

Isolation of Salmonella enterica Serovar Enteritidis from Houseflies (Musca domestica) Found in Rooms Containing Salmonella Serovar Enteritidis-Challenged Hens

ABSTRACT Houseflies (Musca domestica) released into rooms containing hens challenged with Salmonella enterica serovar Enteritidis (Salmonella serovar Enteritidis) rapidly became contaminated with Salmonella serovar Enteritidis. Forty to 50% of the flies were contaminated at 48 h, and the percentage increased to 50 to 70% at 4 and 7 days postexposure and then decreased to 30% at day 15. Initial attempts at recovering surface organisms for culture using an aqueous rinse were largely unsuccessful, while cultures of internal contents readily recovered Salmonella serovar Enteritidis. However, when 0.5% detergent was incorporated into the rinse, high recovery levels of bacteria were observed from both external and internal culture regimens, indicating equal distribution of the organism on and in the fly and a tighter interaction of the organism with the host than previously thought. Salmonella serovar Enteritidis was isolated routinely from the fly gut, on rare occasions from the crop, and never from the salivary gland. Feeding contaminated flies to hens resulted in gut colonization of a third of the birds, but release of contaminated flies in a room containing previously unchallenged hens failed to result in colonization of any of the subject birds. These results indicate that flies exposed to an environment containing Salmonella serovar Enteritidis can become colonized with the organism and might serve as a source for transmission of Salmonella serovar Enteritidis within a flock situation.

Nicotinoid and pyrethroid insecticide resistance in houseflies (Diptera: Muscidae) collected from Florida dairies

BACKGROUND The housefly, Musca domestica L., continues to be a major pest of confined livestock operations. Houseflies have developed resistance to most chemical classes, and new chemistries for use in animal agriculture are increasingly slow to emerge. Five adult housefly strains from four Florida dairy farms were evaluated for resistance to four insecticides (beta-cyfluthrin, permethrin, imidacloprid and nithiazine). RESULTS Significant levels of tolerance were found in most field strains to all insecticides, and in some cases substantial resistance was apparent (as deduced from comparison with prior published results). At the LC(90) level, greater than 20-fold resistance was found in two of the fly strains for permethrin and one fly strain for imidacloprid. Beta-cyfluthrin LC(90) resistance ratios exceeded tenfold resistance in three fly strains. The relatively underutilized insecticide nithiazine had the lowest resistance ratios; however, fourfold LC(90) resistance was observed in one southern Florida fly strain. Farm insecticide use and its impact on resistance selection in Florida housefly populations are discussed. CONCLUSION Housefly resistance to pyrethroids is widespread in Florida. Imidacloprid resistance is emerging, and tolerance was observed to both imidacloprid and nithiazine. If these insecticides are to retain efficacy, producer use must be restrained.

Effect of Habitat Depth on Host Location by Five Species of Parasitoids (Hymenoptera: Pteromalidae, Chalcididae) of House Flies (Diptera: Muscidae) in Three Types of Substrates

Abstract Four species of pteromalid parasitoids [Muscidifurax raptor Girault & Sanders, Spalangia cameroni Perkins, Spalangia endius Walker, Spalangia gemina Boucek, and the chalcidid Dirhinus himalayanus (Masi)] were evaluated for their ability to locate house fly pupae at various depths in poultry manure (41% moisture), fly rearing medium (43% moisture), and sandy soil (4% moisture) from a dairy farm. Searching activity in manure was largely confined to the surface (M. raptor, D. himalayanus, and S. gemina) or to depths of up to 2 cm below the surface (S. endius, S. cameroni). S. cameroni was the most effective species at locating buried pupae in manure. All of the species searched over a wider range of habitat depths in fly rearing medium, although M. raptor and S. gemina tended to concentrate their searching activity relatively close to the surface of the substrate. Host attacks by these species at 6 cm were 30–40% lower than on the surface of the medium. S. endius searched uniformly at all depths in rearing medium and S. cameroni had highest rates of host attacks 1–2 cm below the surface of this substrate. The parasitoids displayed considerable fidelity to their search patterns regardless of whether or not they were given a choice of habitat depths in which they could find pupae. None of the parasitoids were effective at attacking fly pupae that were buried in sandy soil at any depth. The results suggest that fly larvae that pupate in the sandy soils typical of Florida’s coastal plains are relatively impervious to attack by pupal parasitoids.

Suppression of House Flies (Diptera: Muscidae) in Florida Poultry Houses by Sustained Releases of Muscidifurax raptorellus and Spalangia cameroni (Hymenoptera: Pteromalidae)

Abstract Weekly releases of Muscidifurax raptorellus Kogan and Legner and Spalangia cameroni Perkins were made for 12 wk after house cleanouts in Florida pullet houses in either spring/summer (May–August) or fall (September–December). Releases were made by weekly placement of 62,500 and 85,000 pupae parasitized by M. raptorellus and S. cameroni, respectively, which produced an average of 79,049 and 32,841 adult female parasitoids per week. House fly (Musca domestica L.) pupal mortality, as measured by sentinel pupae, was about twice as high in the release house (40.2%) as in the two control houses (21.5 and 21.8%) in the summer release. Pupal mortality in the fall was three to four times higher in the release house (45.6%) as in the two control houses (13.6 and 8.4%). Although successful parasitism of sentinel pupae was only ≈8.4% in the release houses in both studies, parasitism was significantly higher than the control houses in both summer (3.9 and 1.7%) and fall (0.0 and 0.8%) releases. Fly populations were high in both studies but significantly lower in the release houses than the controls in both summer (361.5 versus 450.3 and 584.4 spots/spot card/wk) and fall (477.1 versus 971.4 and 851.8 spots/card/wk) releases. An average of 4.8 M. raptorellus emerged from each pupa parasitized by this species, with parasite loads ranging from 1 (8.6%) to 17 (0.07%) adults emerged per parasitized pupa.

Gas chromatography/mass spectrometry analysis of the cuticular hydrocarbons from parasitic wasps of the genus Muscidifurax

Parasitic Hymenoptera can be difficult to identify by conventional taxonomic techniques. Examination of the cuticular hydrocarbons (CHCs) provides a basis for chemotaxonomic differentiation, which may lead to the discovery of pheromones, and can be a means of examining colonies for species cross-contamination. The parasitic wasps examined were Muscidifurax raptor, M. zaraptor, M. uniraptor, and the gregarious form of M. raptorellus. Species within the genus Muscidifurax, as well as the sex, can clearly be differentiated by examining the gas chromatograms of the CHCs. Identification of the alkanes by mass spectrometry shows uncommon dimethylalkanes and trimethylalkanes for members of the genus. The methyl branched cuticular hydrocarbons of these insects are rare compared to those found on insects reported in the literature, but are present in significant amounts on these insects. Additionally, sexual dimorphism is observed in long chain alkanes (C21–C39) present on male and female cuticular surfaces for these species. Females tend to have cuticular hydrocarbons with methyl branches located externally on the carbon backbone chain for dimethyl-, trimethyl-, and tetramethylalkanes, whereas males tend to have dimethyl- and trimethylalkanes located internally on the hydrocarbon backbone chains. Mass spectra of novel and rare methyl branched compounds identified on these parasitoids are presented.

Effects of Salivary Gland Hypertrophy Virus on the Reproductive Behavior of the Housefly, Musca domestica

ABSTRACT Pathological studies demonstrated that the salivary gland hypertrophy virus of houseflies (MdSGHV) shuts down reproduction in infected females. The mechanism that underlay the disruption of reproduction functioned on several levels. Females infected at the previtellogenic stage did not produce eggs, reflecting a block in the gonadotropic cycle. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot analysis of hemolymph samples demonstrated that MdSGHV infection reduced the levels of both the female-specific hexamerin and egg yolk proteins. Furthermore, reverse transcriptase quantitative real-time PCR data demonstrated that infection blocked hexamerin and yolk protein gene transcription. When females were allowed to develop eggs prior to infection (postvitellogenic stage), the outcome of mating attempts depended upon when mating took place. If egg-containing, virus-infected females were mated within 24 h of infection, they copulated and deposited a single batch of fertilized eggs. However, if mating was delayed for a longer period, the egg-containing females refused to copulate with healthy males. Both of these results suggested that a virus-induced signal influenced the central nervous system, shutting down female receptivity and egg production. All experiments demonstrated that MdSGHV-infected males did not display azoospermia and were fertile. Both healthy females mated with infected males, and the resulting F1 progeny were free of salivary gland hypertrophy symptoms, which suggests that the virus is not sexually or vertically transmitted.

Insecticide resistance in house flies from the United States: Resistance levels and frequency of pyrethroid resistance alleles

Although insecticide resistance is a widespread problem for most insect pests, frequently the assessment of resistance occurs over a limited geographic range. Herein, we report the first widespread survey of insecticide resistance in the USA ever undertaken for the house fly, Musca domestica, a major pest in animal production facilities. The levels of resistance to six different insecticides were determined (using discriminating concentration bioassays) in 10 collections of house flies from dairies in nine different states. In addition, the frequencies of Vssc and CYP6D1 alleles that confer resistance to pyrethroid insecticides were determined for each fly population. Levels of resistance to the six insecticides varied among states and insecticides. Resistance to permethrin was highest overall and most consistent across the states. Resistance to methomyl was relatively consistent, with 65-91% survival in nine of the ten collections. In contrast, resistance to cyfluthrin and pyrethrins + piperonyl butoxide varied considerably (2.9-76% survival). Resistance to imidacloprid was overall modest and showed no signs of increasing relative to collections made in 2004, despite increasing use of this insecticide. The frequency of Vssc alleles that confer pyrethroid resistance was variable between locations. The highest frequencies of kdr, kdr-his and super-kdr were found in Minnesota, North Carolina and Kansas, respectively. In contrast, the New Mexico population had the highest frequency (0.67) of the susceptible allele. The implications of these results to resistance management and to the understanding of the evolution of insecticide resistance are discussed.

Salivary Gland Hypertrophy Viruses: A Novel Group of Insect Pathogenic Viruses

Salivary gland hypertrophy viruses (SGHVs) are a unique, unclassified group of entomopathogenic, double-stranded DNA viruses that have been reported from three genera of Diptera. These viruses replicate in nuclei of salivary gland cells in adult flies, inducing gland enlargement with little obvious external disease symptoms. Viral infection inhibits reproduction by suppressing vitellogenesis, causing testicular aberrations, and/or disrupting mating behavior. Historical and present research findings support a recent proposal of a new virus family, the Hytrosaviridae. This review describes the discovery and prevalence of different SGHVs, summarizes their biochemical characterization and taxonomy, compares morphological and histopathological properties, and details transmission routes and the influence of infection on host biology and reproduction. In addition, the potential use of SGHVs as sterilizing agents for house fly control and the deleterious impact of SGHVs on colonized tsetse flies reared for sterile insect technique are discussed.

Microsporidioses of Muscidifurax raptor (Hymenoptera:Pteromalidae) and other pteromalid parasitoids of muscoid flies

Abstract An undetermined microsporidium was found infecting Muscidifurax raptor, a parasitoid of muscoid flies. Housefly hosts of the parasitoid in culture were not infected. Spores were present in newly deposited parasitoid eggs and vertical transmission experiments indicated 100% efficiency of maternal transmission. An uninfected culture was established by isolating parasitoids for oviposition and discarding the progeny of infected females. Infected parasitoids took longer to complete development, lived half as long, and produced 1 8 to 1 2 as many progeny as uninfected parasitoids. Thirty-five percent of field-collected parasitoids from New York dairy farms in August of 1990 were infected, although infection increased to 100% after two generations in culture. A survey of 11 pteromalid species collected from different geographic areas indicated that microsporidioses in Muscidifurax raptor and M. zaraptor are ubiquitous. Urolepis rufipes was the only other species examined that was also infected with microsporidia.

Selection for Resistance to Imidacloprid in the House Fly (Diptera: Muscidae)

ABSTRACT The house fly, Musca domestica L. (Diptera: Muscidae), continues to be a primary pest of livestock facilities worldwide. This pest also has shown a propensity for pesticide resistance development when under high selection pressures. In this study the house fly strain FDm was created by a 20% contribution from each of five colonies collected from dairies in Florida with known imidacloprid resistance. The FDm strain was used to evaluate the level of imidacloprid resistance after five selections near the LC70 value of each selected generation. Overall, the mean selection mortality was 72.7, with males being considerably more susceptible than females. The unselected (F0) FDm strain showed considerable susceptibility to imidacloprid after its creation, compared with the five parental strains. Between 9,500 and 14,000 virgin house flies were used in each selection. After the fifth and final selection, a 331-fold increase in imidacloprid resistance at the LC70 was observed over the parental FDm strain. In parallel studies, the FDm strain showed increasing tolerance of the commercial imidacloprid product QuickBayt. These results suggest that livestock producers should use caution when choosing pesticides and consider rotating fly baits, as is encouraged with other pesticide treatment regimes on farms.