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IntroductionNosema is a diverse genus of unicellular microsporidian parasites of insects and other arthropods. Nosema muscidifuracis infects parasitoid wasp species of Muscidifurax zaraptor and M. raptor (Hymenoptera: Pteromalidae), causing ~50% reduction in longevity and ~90% reduction in fecundity.Methods and ResultsHere, we report the first assembly of the N. muscidifuracis genome (14,397,169 bp in 28 contigs) of high continuity (contig N50 544.3 Kb) and completeness (BUSCO score 97.0%). A total of 2,782 protein-coding genes were annotated, with 66.2% of the genes having two copies and 24.0% of genes having three copies. These duplicated genes are highly similar, with a sequence identity of 99.3%. The complex pattern suggests extensive gene duplications and rearrangements across the genome. We annotated 57 rDNA loci, which are highly GC-rich (37%) in a GC-poor genome (25% genome average). Nosema-specific qPCR primer sets were designed based on 18S rDNA annotation as a diagnostic tool to determine its titer in host samples. We discovered high Nosema titers in Nosema-cured M. raptor and M. zaraptor using heat treatment in 2017 and 2019, suggesting that the remedy did not completely eliminate the Nosema infection. Cytogenetic analyses revealed heavy infections of N. muscidifuracis within the ovaries of M. raptor and M. zaraptor, consistent with the titer determined by qPCR and suggesting a heritable component of infection and per ovum vertical transmission.DiscussionThe parasitoids-Nosema system is laboratory tractable and, therefore, can serve as a model to inform future genome manipulations of Nosema-host system for investigations of Nosemosis.

Entomopathogenic fungi such as Beauveria bassiana (Balsamo) Vuillemin and Metarhizium anisopliae/brunneum (Metchnikoff)/Petch have shown promising results for managing the house fly, Musca domestica L. A primary challenge of using these biological control agents (BCAs) in field situations is the time required to induce high adult house fly mortality, typically 6–7 d post-exposure. In this study, virulence of M. anisopliae (strain F52) and four B. bassiana strains were compared. The B. bassiana strains GHA and HF23 are used in commercial products and those were compared with two strains that were isolated from house flies on dairy farms (NFH10 and L90). Assays were conducted by exposing adult house flies to fungal-treated filter paper disks for 2 h. The lethal time to 50% mortality (LT50) at the high concentration of 1 × 109 conidia ranged from 3.8 to 5.2 d for all five strains. GHA, NFH10, and L90 killed flies faster than M. anisopliae strain F52; HF23 did not differ from either the M. anisopliae or the other B. bassiana strains. Attempts with the NFH10 strain to induce faster fly mortality through selection across 10 fungal to fly passages did not result in shorter time to fly death of the selected strain compared with the unselected strain.

Insects are ectothermic organisms; hence, all aspects of their biology are strongly influenced by ambient temperatures. Different insect species respond differently with phenotypic plasticity and/or genetic adaptation to changing temperatures. Here, we tested the thermal adaptation of the house fly and three of its parasitoids species by comparing life-history parameters in populations from a hot climate region (Jordan Valley) and from a moderate-climate region (Galilee). No significant differences were found between the two house fly populations, both under hot and moderate experimental conditions. Life-history parameters of the parasitoids (Muscidifurax raptor Girault & Sanders, Spalangia endius Walker, and Spalangia cameroni Perkins [Hymenoptera: Pteromalidae]) varied markedly between origins, species, sexes, and experimental conditions. Of the three species tested, only M. raptor collected in the Jordan Valley proved better adapted to experimental heat conditions, compared to its counterpart population that was collected in the Galilee. Additionally, we tested the effect of elevating temperatures on a house fly lab population for 17 consecutive generations and found no evidence for heat adaptation. We discuss our results in the context of house fly control and global warming.

The house fly, Musca domestica L. (Diptera: Muscidae), is a global pest of humans and animals that carries scores of pathogens and costs up to $1 billion per year in the United States alone. Information is reviewed on recognition, distribution, biology, dispersal, and associations with microbes. Particular challenges of managing flies in different animal systems are discussed for swine, poultry, dairy cattle, beef feedlot, and equine operations. Effective fly management requires diligent monitoring and integration of cultural control, especially manure management, with mechanical control, traps, conservation or augmentative biological control, and judicious use of insecticides. House fly is notorious for developing insecticide resistance and its resistance status is summarized as of August 2020. Several critical research needs are identified. Monitoring systems and nuisance/action thresholds need improvement. Faster-killing strains and better formulations are needed to integrate pathogens into Integrated Pest management (IPM) programs. The use of parasitoids remains an inexact science with many questions remaining about species selection and release rates. New attractants are needed for use in traps and attract-and-infect/kill strategies. Screening of new active ingredients for toxicity should continue, including a rigorous assessment of essential oils and other botanicals. Rising global temperatures may affect the balance of the fly with natural enemies. An understanding of the fly microbiome may reveal unknown vulnerabilities, and much remains to be learned about how flies acquire, retain, and transmit human and animal pathogens. System-specific research is also needed to tailor fly IPM programs to individual animal systems, especially in organic and free-range animal production.

Veterinary and medical entomologists who are involved in research on pest control often need to perform dose–response bioassays and analyze the results. This article is meant as a beginner’s guide for doing this and includes instructions for using the free program R for the analyses. The bioassays and analyses are described using previously unpublished data from bioassays on house flies, Musca domestica Linnaeus (Diptera: Muscidae), but can be used on a wide range of pest species. Flies were exposed topically to beta-cyfluthrin, a pyrethroid, or exposed to spinosad or spinetoram in sugar to encourage consumption. LD50 values for beta-cyfluthrin in a susceptible strain were similar regardless of whether mortality was assessed at 24 or 48 h, consistent with it being a relatively quick-acting insecticide. Based on LC50 values, spinetoram was about twice as toxic as spinosad in a susceptible strain, suggesting a benefit to formulating spinetoram for house fly control, although spinetoram was no more toxic than spinosad for a pyrethroid-resistant strain. Results were consistent with previous reports of spinosad exhibiting little cross-resistance. For both spinosad and spinetoram, LC50 values were not greatly different between the pyrethroid-resistant strain and the susceptible strain.

Filth flies remain one of the most prevalent pest groups affecting the animal production industry. Spalangia spp. and Muscidifurax spp. are beneficial parasitic wasps that often are utilized to manage filth fly populations such as house flies, Musca domestica L. (Diptera: Muscidae), and stable flies, Stomoxys calcitrans (L.) (Diptera: Muscidae). These wasps search for filth fly pupae as hosts in areas potentially treated with insecticides, which may result in nontarget insecticide selection effects. However, research regarding resistance development in parasitic wasps such as S. cameroni Perkins (Hymenoptera: Pteromalidae) is limited. Therefore, a study was conducted to determine the potential of S. cameroni to develop resistance to the commonly used insecticide permethrin, as well as compare permethrin susceptibility among several S. cameroni strains. After 10 selected generations, susceptibility was significantly lower for the selected strain when compared with that of its unselected parent strain. A comparison of several parasitoid strains collected from different U.S. states indicated that permethrin susceptibility was not significantly different between a baseline strain and more recently established field strains. The potential implications of this previously unrecognized nontarget insecticide exposure effect on filth fly parasitoids are discussed.

House fly (Musca domestica) control is a major challenge in animal agriculture. Here, we tested the feasibility of applying pyriproxyfen (PPF), an insect-growth regulator that controls house flies effectively, using autodissemination methods, in which the flies themselves deliver PPF to their oviposition sites. First, we tried baiting gravid female flies to walk-through stations, where flies would self-treat with PPF and distribute it. This concept worked well in laboratory and indoor cage experiments, but not in the field, as flies appeared reluctant to alight on and collect PPF. Therefore, we tested a different concept of actively coating flies with PPF and then releasing them in different proportions. This concept was tested in laboratory experiments with various manure types in the USA and in Israel. Twenty percent of PPF-coated flies (corresponding to ≥ 2.3 mg/kg PPF) were sufficient to get high control levels (~ 90%) in most of the tested manure types in the US study. Very similar results were obtained in the experiments in Israel but only with poultry manure, whereas low control levels were obtained when cow manure was used. We conclude that autodissemination of PPF using the collect–treat–release “active coating” concept may be practical, depending on manure type, and should be further tested in the field.

Filth flies cause billions of dollars of losses annually to the animal production industry. Fluralaner is a relatively new pesticide currently sold for control of fleas, ticks, and mites on companion animals and poultry. We examined the efficacy of fluralaner against three species of filth flies. Insecticide-susceptible horn flies and stable flies were tested topically. Fluralaner outperformed permethrin by > 2-fold for the horn flies but underperformed permethrin by > 45-fold for stable flies at 24 h. House flies were tested topically with fluralaner in comparison to permethrin at 48 h and orally with fluralaner in comparison to imidacloprid at 24 h. Topical fluralaner was 6- to 28-fold as toxic as permethrin in four pyrethroid-resistant strains and not significantly less toxic than permethrin in a susceptible strain and a mildly pyrethroid-resistant strain. There was slight cross-resistance between topically applied fluralaner and permethrin in all five insecticide-resistant strains tested. Oral fluralaner was more toxic than imidacloprid in all four house fly strains tested, 9- to 118-fold as toxic. Oral cross-resistance between imidacloprid and fluralaner was not detected, but imidacloprid resistance was not high in any of the tested strains. Fluralaner shows promise for control of horn flies and house flies.

Lucilia sericata is a facultative ectoparasite causing fly strike or myiasis in warm-blooded vertebrates. It is controlled by traps or insecticides, but both have drawbacks and alternative ways of control are urgently needed. Essential oils (EOs) of vetiver (Chrysopogon zizanioides), cinnamon (Cinnamomum zeylanicum), and lavender (Lavandula angustifolia) and their blends (OBs); OB1 (2 ml of each EO plus 4 ml of sunflower oil as a carrier) and OB2 (2 ml of each EO) were tested. Oils were tested at 5% for deterrence assays, and a dose response assay 0.01-0.6%, was conducted to determine forced-contact toxicity. We evaluated the efficacy of oils as oviposition deterrents, repellents/attractants, and their effects on mortality and longevity of adult L. sericata. Our data indicated that 0.2% EOs killed all flies by 5 min post-treatment and that vetiver oil greatly deterred flies from the oviposition medium and reduced adult longevity. Sunflower oil repelled all flies from ovipositing and greatly reduced the lifespan of treated adults. The blend of the four oils (OB1) had the greatest repellent effect on the flies. EOs have insecticidal, repellent, and oviposition-deterrent activities against L. sericata that could be used for suppression of blow fly populations.

The pteromalid pupal parasitoids Muscidifurax raptor Girault & Sanders and Spalangia cameroni Perkins (Hymenoptera: Pteromalidae) are commonly released on livestock farms for management of house flies, Musca domestica L. (Diptera: Muscidae). To be effective, parasitoids must be able to locate live host puparia in complex environments that may include dead or formerly parasitized hosts and non-host physical objects. In this study, both species of parasitoids were examined for their ability to kill and parasitize live house fly puparia either alone or in mixtures with formerly parasitized (dead) hosts or similarly sized acrylic beads. Muscidifurax raptor killed significantly fewer hosts and produced fewer progeny when the parasitoids were provided with hosts that were mixed with formerly parasitized puparia. Spalangia cameroni was unaffected by the presence of formerly parasitized puparia for any of the measured variables. When beads were used as a decoy instead of formerly parasitized puparia, high bead-to-live-host ratios (90% decoys) resulted in significantly fewer numbers of hosts killed by M. raptor compared with the other treatments (50% and no decoys). Residual host mortality at the high bead-to-live-host ratio (90% decoys) was lower (31.2%) than in ratios of 50:50 and with no decoys (51.6 and 59.3%, respectively), so that progeny production by M. raptor was unaffected by the presence of beads. Spalangia cameroni killed over twice as many hosts and produced twice as many progeny in the absence of bead decoys than when beads made up 90% of the decoy—host mixture. The results support the scatter method for deploying parasitized puparia during releases, because the presence of formerly parasitized hosts did not interfere substantially with the ability of S. cameroni and M. raptor to locate and parasitize live pupae.

House flies, Musca domestica L., are widely recognized for their ability to develop resistance to chemical insecticides so alternative control strategies are desired. The use of entomopathogenic fungi such as Beauveria bassiana (Balsamo) Vuillemin to manage house fly populations has shown promising results; however, the success of using this fungus against larval house flies varies widely. The overall objective of this study was to examine factors that may influence efficacy of B. bassiana treatments against larvae. When a high concentration (4 × 1011 conidia/ml) was applied to first- and second-instar larvae in rearing medium, there was a significant reduction in pupation and adult emergence rates. Treating third-instar larvae at the same concentration did not result in a significant reduction of pupation or adult emergence. Temperature (22 versus 32°C) and media composition (diets with- and without propionic acid) did not affect the B. bassiana treatment efficacy against house fly larvae. The narrow time window of vulnerability of larvae and the high doses required to infect them indicate that B. bassiana has little potential as an operational biocontrol agent for house fly larvae.

Parasitoids are important natural enemies of house flies and other muscoid flies. The two most commonly used methods for collecting fly parasitoids from the field have distinct advantages and disadvantages. Collections of wild puparia depend on the ability to find puparia in sufficient numbers and are prone to localized distortions in relative species abundance because of the overrepresentation of samples from hot spots of fly larval activity. Placement and retrieval of sentinel puparia is convenient and allows consistent sampling over time but is strongly biased in favor of Muscidifurax spp. over Spalangia spp. An improved sentinel method is described that combines some of the advantages of these two methods. Fly medium containing larvae is placed in containers, topped with a screen mesh bag of puparia, and placed in vertebrate-proof wire cages. Cages are placed at sites of actual or potential fly breeding and retrieved 3–7 d later. The modified method collected species profiles that more closely resembled those of collections of wild puparia than those from sentinel pupal bags. A method is also described for isolating puparia individually in 96-well tissue culture plates for parasitoid emergence. Use of the plate method provided a substantial saving of time and labor over the use of individual gelatin capsules for pupal isolation. Puparia from the collections that were housed individually in the wells of tissue culture plates had a higher proportion of emerged Spalangia species than puparia that were held in groups.

The entomopathogenic fungus Beauveria bassiana (Balsamo) Vuillemin (Hypocreales: Cordycipitaceae) has been widely studied against a wide range of arthropod pests, including many of medical and veterinary importance. New investigators must sort through a wide array of published methods for the production, harvest, storage, and bioassay methods for this pathogen. Simplified methods for production of conidia using Sabouraud dextrose agar with yeast (SDYA) plates and two conidial harvesting methods are described. Dry harvesting yields conidia that are ready to incorporate into dusts and food baits, but the fungal product includes mycelial debris that can hamper quantification and introduces variable amounts of unwanted bulk. Wet harvesting with filtration produces a cleaner product that is immediately ready for testing in liquid formulations. Examples of bioassays with house flies are presented that include conidia applied topically to the dorsal thorax for dose–mortality assays and conidial suspensions applied to filter paper disks for concentration mortality assays.

House flies are global pests and notoriously difficult to control. Essential oils of vetiver, cinnamon, and lavender and their blends were tested for toxic and repellent effects against larval and adult flies. All of the oils had moderate toxicity for eggs. Mortality of 2nd instar larvae was 57-78% in dipping assays, 38-100% in contact assays, and 94-100% in treated media. Lavender was less effective (38% mortality) than the others (91-100%) in contact bioassays. Oil blends were not more effective against larvae than individual oils. Vetiver and cinnamon oils were strongly repellent (84 and 78%, respectively) for larvae in treated media. None of the oils were repellent for adult house flies in olfactometer assays, but testing of additional products demonstrated significant repellency for neem oil, p-menthane-3,8-diol (PMD), and vanillin. Contact/fumigant toxicity of vetiver, cinnamon, and lavender oils was 100%, significantly higher than mortality from sunflower oil (67%). Blends of oils were not more effective against adults than the individual oils, but blends diluted with sunflower oil were as effective as the individual oils. Essentials oils of vetiver and cinnamon may have potential for fly management in situations where conventional insecticides cannot be used.

Colonies of house flies (Musca domestica L. [Diptera: Muscidae]) and four species of parasitoids (Muscidifurax raptor Girault and Sanders, Muscidifurax zaraptor Kogan and Legner, Spalangia cameroni Perkins and Spalangia endius Walker) were established by making collections from dairy farms near Bell, FL, Beatrice, NE, Minneapolis, MN, and San Jacinto, CA. Colonies were assessed for heat tolerance by comparing life history parameters at 25–27°C and fluctuating hot (26.7–41.7°C) temperatures. Muscidifurax raptor, S. cameroni, and S. endius produced 24–28% as many progeny under hot conditions as at 25°C. Colonies of M. zaraptor were more heat-tolerant and produced an average 46.9% as many progeny under the hot regime compared with moderate conditions. There was little evidence for higher heat tolerance in parasitoid populations from historically hot locations (CA desert and FL). Colonies of M. raptor and S. endius that had been in culture for 24 yr were the least heat-tolerant with regard to progeny production. House flies collected from the same locations varied little in longevity, fecundity, or egg-to-adult survival under either hot or moderate regimes. Flies reared under hot conditions laid about half as many eggs (89/female) and had about half the egg–adult survival rate (47.3%) under hot compared with moderate conditions, indicating that heat stress had less effect on flies than on all of the parasitoids except M. zaraptor. An attempt to select for heat tolerance in flies by subjecting them to incremental increases in rearing temperatures for 20 generations resulted in little change in tolerance among the selected flies.

Synthetic dsRNAs, used to induce RNA interference, may have dose dependent phenotypic effects. These effects are difficult to define if the dsRNAs are delivered using a non-quantitative method. Accurate delivery of known quantities of nucleic acids or other chemicals is critical to measure the efficacy of the compound being tested and to allow reliable comparison between compounds. Here we provide a reproducible, quantitative microinjection protocol that ensures accurate delivery of specific doses of dsRNA, reducing the mortality typically induced by injection injury. These modifications include the addition of Rhodamine B, a graduated injection needle, and an improved recovery method borrowed from Isoe and Collins. This method allows calculation of dose responses and facilitates comparisons between compounds. Versions of this method have been successfully used on three genera of mosquitoes as well as house flies to assess the reduction in fecundity resulting from gene silencing of ribosomal RNA transcripts. This protocol provides strategies to reduce several challenges of small insect microinjection. Together, mechanical delivery of dsRNAs accompanied by visual verification, identification of effective locations for delivery, and inclusion of a post-injection recovery period ensure accurate dosing and low injury mortality. This protocol also describes an oviposition bioassay for uniform determination of effects on fecundity.

The house fly, Musca domestica (L.) (Diptera: Muscidae), and the stable fly, Stomoxys calcitrans (L.) (Diptera: Muscidae), are two filth flies responsible for significant economic losses in animal production. Although some chemical control products target adults of both species, differences in mouthpart morphology and behavior necessitates distinct modalities for each. For these reasons, larvicides are an attractive means of chemical control. We assessed the potential of the polyol sweeteners erythritol and xylitol as larvicides to the house fly and stable fly. LC50 values of erythritol against 2nd instar larvae were 34.94 mg/g media (house fly) and 22.10 mg/g media (stable fly). For xylitol, LC50 values were 74.91 mg/g media (house fly) and 41.58 mg/g media (stable fly). When given a choice, neither species showed a preference for ovipositing in media treated with either sweetener at various concentrations or in media without sweetener. Significantly lower development from egg to adult was observed when the 2nd instar LC50 equivalent of each sweetener was present in the media compared to controls. Erythritol and xylitol both have larvicidal qualities, however their effective concentrations would necessitate creative product formulation and deployment methods to control all stages of developing flies.

Interest in biological methods for livestock and poultry pest management is largely motivated by the development of resistance to most of the available synthetic pesticides by the major pests. There also has been a marked increase in organic systems, and those that promote animal welfare by reducing animal densities and allowing greater freedom of movement. Such systems, especially organic operations, are in need of new tools and strategies to manage pest problems. This chapter reviews the status of entomopathogenic fungi, viruses, and nematodes for management of pests of livestock and poultry production. The pests covered here are limited to the research that has been conducted to date, but include biting and nuisance flies, ticks, poultry ectoparasites, and litter beetles. Many of the same pests affect horses as well, although little work has been done in this area. The fungal pathogens Beauveria bassiana and Metarhizium anisopliae s.l. have received the most attention and have been evaluated against most of the major livestock and poultry pests. These pathogens have the greatest potential for practical use, but more work is needed to identify efficacious isolates and develop formulations for both on- and off-host use. The slow kill rate of these pathogens has been considered a liability in the past, but a growing body of work has demonstrated important sublethal effects of infection that includes reduced feeding, movement, and pathogen transmission. Two pathogens of house fly, the fungus Entomophthora muscae and salivary gland hypertrophy virus, are effective under some conditions but have biological characteristics that have limited their development as practical management tools. Entomopathogenic nematodes (Steinernema and Heterorhabditis spp.) also have potential but are limited by environmental constraints, especially temperature and substrate moisture. Discovery of, or selection for, strains with wider environmental tolerance could broaden the range of situations where they can be used effectively. © Wageningen Academic Publishers 2018.

The pupal parasitoids, Spalangia cameroni Perkins and Muscidifurax raptor Girault and Sanders, can be purchased for biological control of house flies Musca domestica L. and stable flies Stomoxys calcitrans (L.) (Diptera: Muscidae). Little is known about the odors involved in host-seeking behavior of these two species, so odors associated with house flies were investigated in the laboratory using a Y-tube olfactometer. Odor stimuli from house fly host puparia, larvae, pine-shavings bedding with horse manure, and developing flies in the pine-shavings-manure substrate were evaluated in bioassays using the two pteromalid species. In choice tests, naïve female S. cameroni were strongly attracted to odor from the substrate containing house fly larvae and secondarily from the uninfested substrate and substrate with puparia versus humidified and purified air. This species also selected the substrate with larvae versus the substrate with the house fly puparia or uninfested substrate. Muscidifurax raptor was attracted to odor from the substrate containing puparia, washed puparia, and substrate with puparia removed. The data suggest that coexistence between the two pteromalid parasitoids, S. cameroni and M. raptor, might be promoted by different host-seeking behavior.

The horn fly, Haematobia irritans (L.) is an important cattle pest and traditionally has been managed using insecticides; however, many horn fly populations are insecticide-resistant in United States. Use of alternative control techniques has been limited because of the challenges of managing a fly pest on pastured cattle. After the discovery of a wild horn fly infected with Beauveria bassiana in Florida, the fungus was cultured and evaluated for efficacy against laboratory-reared horn flies. This fungal strain was selected for increased virulence by passage through laboratory-reared horn fly hosts to shorten interval from infection to fly death and subsequent conidia formation, properties important to future use of the fungus as a biological control agent against horn flies. After seven passages through horn fly hosts, fly mortality was not significantly accelerated as evaluated through LT50 values, but conidia were readily produced from these killed flies. Although further development is needed to improve fungal efficacy, this fungal strain holds promise as a biological control agent for inclusion in horn fly integrated pest management programs.

Competition bioassays were conducted with the filth fly pupal parasitoids Muscidurax raptor (Girault & Sanders) and M. raptorellus (Kogan & Legner) (Hymenoptera: Pteromalidae) using house fly Musca domestica L. (Diptera: Muscidae) hosts at different host densities. Muscidifurax raptor had a significant impact on M. raptorellus when hosts were limiting in sequential parasitism tests. Fewer than six M. raptorellus adult progeny emerged from groups of 50 fly pupae that were parasitized by M. raptor at the same time or when M. raptor parasitism preceded M. raptorellus by 48 h, respectively, compared with 42–55 M. raptorellus progeny produced when this species was tested alone. Production of M. raptor was significantly lower when parasitism by this species was preceded by M. raptorellus (25) than when M. raptor was tested alone (43). When the two species parasitized hosts at the same time in different proportions at low host:parasitoid densities (5:1), M. raptorellus produced 13 progeny per parent female when it was the sole species present and fewer than two when M. raptor was present. No negative impact of M. raptorellus on M. raptor was observed. Neither species had a substantial effect on the success of the other at higher host:parasitoid densities.

House flies, Musca domestica L., and stable flies, Stomoxys calcitrans (L.), (Diptera: Muscidae), common pests on equine facilities, were studied in the laboratory to determine the success and duration of larval development and oviposition preferences on six substrates commonly found on equine facilities. Substrates tested were hay soiled with urine and manure, fresh horse manure, pine shaving bedding soiled with urine and manure (12 h old), pine shaving bedding soiled with urine and manure (aged72 h in a manure pile), builders sand bedding soiled with urine and manure aged 3 d, and soil from an overgrazed pasture mixed with urine and manure of variable age. House fly larvae failed to develop into adults in hay, soil, and sand substrates. Stable flies preferred to oviposit on substrates with plant material and not on fresh manure. However, when eggs were added to the substrates, pupariation was maximal in fresh manure and the fresh pine shaving substrate. Stable flies developed in all six equine substrates, but development was less successful on the substrates with soil. In choice tests, fresh manure and the fresh pine shaving substrates were the most attractive for house fly oviposition. These substrates also yielded the greatest number of house fly puparia from artificially added eggs. An understanding of oviposition preferences and differential larval development of house flies and stable flies on these substrates may help develop options for reducing pest populations by effectively managing equine waste and selecting appropriate bedding materials.

In this paper it is proposed that one potential component by which the Musca domestica salivary gland hypertrophy virus (MdSGHV) infects individual flies is through cuticular damage. Breaks in the cuticle allow entry of the virus into the hemocoel causing the infection. Male flies typically have a higher rate of infection and a higher rate of cuticular damage than females. A model for the transmission of MdSGHV was formulated assuming several potential and recognized means of transmission. The model yields results that are in agreement with field data that measured the infection rate in house flies on dairy farms in Florida. The results from this model indicate that MdSGHV will be maintained at a stable rate within house fly populations and support the future use of MdSGHV as a birth control agent in house fly management.

House flies, (Musca domestica L.) and stable flies (Stomoxys calcitrans L.) are common pests on equine facilities. Biological control of these flies with pupal parasitoids has become increasingly popular with horse owners but has not been evaluated on equine facilities. Little is known of the substrate preferences of filth fly parasitoids on equine facilities, but the success of release programs may be affected by microhabitat preferences. Spalangia cameroni Perkins was evaluated for location preferences for parasitization of house fly and stable fly puparia in six substrates commonly found on equine farms in Florida. Substrates were evaluated at 20:1 and 5:1 H:P ratios and during the experiment parasitoids had access to all substrates simultaneously. No differences were observed between filth fly host species in any of the measured parameters: total host mortality, parasitoid progeny production and residual host mortality. Significant effects of H:P ratio on host mortality and residual mortality were found but not on progeny production. While there were significantly more hosts killed in the aged shavings than the fresh shavings at the 20:1 ratio, no differences were observed at the 5:1 ratio. Additionally, no differences were found in progeny production across substrates at the 20:1 ratio, but higher reproductive success was observed in several substrates at the 5:1 ratio. These results demonstrate that S. cameroni had substrate preferences but that these preferences were absent with reduced host density. This parasitoid species appears to be effective at parasitizing hosts in the common equine substrates of Florida.

Pyriproxyfen is an insect growth regulator with juvenile hormone-like activity that has potential uses for dipterans that are difficult to manage with conventional insecticides, such as house flies (Musca domestica L.). The objectives of this study were to determine the efficacy of this insect growth regulator against house flies using variety of delivery systems and target life stages, including an evaluation of the potential for autodissemination by female flies to larval development sites. Adult female house flies exposed to filter paper (3.75% active ingredient) or sugar treated with pyriproxyfen (0.01-0.1%) produced significantly fewer F1 pupae than untreated flies. Adult emergence from pupae was unaffected. In contrast, treatment of larval rearing medium with 0.35 ml/cm2 of a 12 mg pyriproxyfen/liter preparation had no effect on the number of pupae developing from eggs but markedly inhibited adult emergence from those pupae. There was little difference in susceptibility between an insecticide-susceptible and a wild strain of house fly. The LC50 for inhibiting fly emergence of dust formulations in diatomaceous earth incorporating commercial pyriproxyfen products ranged from 8 to 26 mg/liter, with little difference among products. Compared with untreated flies, significantly fewer pupae were produced at concentrations > 0.5% and no adults were produced at concentrations > 0.05% pyriproxyfen. When gravid females were exposed for 1 h to treated fabric (6 mg pyriproxyfen/cm2) and allowed to oviposit in rearing media containing eggs, sufficient pyriproxyfen was autodisseminated to reduce adult emergence from those eggs by > 99%. Intermittent contact with treated fabric over 2 d reduced adult emergence by 63-76%.

Past surveys of feral house fly populations have shown that Musca domestica salivary gland hypertrophy virus (MdSGHV) has a worldwide distribution, with an average prevalence varying between 0.5% and 10%. How this adult-specific virus persists in nature is unknown. In the present study, experiments were conducted to examine short-term transmission efficiency and long-term persistence of symptomatic MdSGHV infections in confined house fly populations. Average rates of disease transmission from virus-infected to healthy flies in small populations of 50 or 100 flies ranged from 3% to 24% and did not vary between three tested geographical strains that originated from different continents. Introduction of an initial proportion of 40% infected flies into fly populations did not result in epizootics. Instead, long-term observations demonstrated that MdSGHV infection levels declined over time, resulting in a 10% infection rate after passing through 10 filial generations. In all experiments, induced disease rates were significantly higher in male flies than in female flies and might be explained by male-specific behaviors that increased contact with viremic flies and/or virus-contaminated surfaces.

The use of pteromalid parasitoids for muscoid fly control is becoming increasingly common. Two species that are often recommended for releases are Spalangia cameroni Perkins and Muscidifurax raptor Girault and Sanders. This study was conducted to determine if the reduced suitability of freeze-killed pupae for Pteromalidae, particularly S. cameroni, is due to freezing, freezing duration, or the type of freezer used. Processing pupae through freezing had a considerably greater negative effect on rearing of S. cameroni than on M. raptor. Although freezing pupae did reduce M. raptor progeny production, there was no effect of length of storage or type of freezer used. Freezing pupae for 5 months resulted in significantly fewer S. cameroni progeny than freezing pupae for 2 weeks. Although not significant, pupae held in a frostfree freezer produced more progeny than those in a non-frostfree freezer. Measurements of weights of parasitized pupae over time indicated that freezing did not result in accelera...

Attack rates, progeny production, sex ratios, and host utilization efficiency of Muscidifurax raptorellus (Kogan and Legner) (Hymenoptera: Pteromalidae), Tachinaephagus zealandicus Ashmead (Hymenoptera: Encyrtidae), and Trichopria nigra (Nees) (Hymenoptera: Diapriidae) were evaluated in laboratory bioassays with five dipteran hosts: house fly (Musca domestica L.), stable fly (Stomoxys calcitrans L.), horn fly (Hematobia irritans L.), black dump fly [Hydrotaea aenescens (Weidemann)] (Diptera: Muscidae), and a flesh fly (Sarcophaga bullata Parker) (Diptera: Sarcophagidae). M. raptorellus killed and successfully parasitized all five host species and produced an average 2.6 parasitoid progeny from each host. Host attack rates were highest on stable fly and lowest on horn fly; there were no differences among hosts in the total number of progeny produced. T. Zealandicus killed larvae of all fly host species in similar numbers, but parasitism was most successful on H. aenescens and S. bullata and least successful on horn fly and house fly hosts. Significantly more parasitoid progeny emerged from S. bullata (10.2 parasitoids per host) than the other hosts; only 2.5 progeny were produced from parasitized horn fly hosts. Most of the killed puparia that produced neither adult flies nor parasitoids (“duds”) contained dead parasitoids; in house fly, stable fly, and horn fly hosts, >30% of these dudded pupae contained adult wasps that failed to eclose. T. nigra successfully parasitized pupae of all host species except house fly and was most successful on stable fly. Significantly more parasitoid progeny emerged from S. bullata (30.6 parasitoids per host) than the other hosts; only 5.7 progeny were produced from horn fly hosts.

The efficiency of host-seeking behavior is crucial to the reproductive performance of female parasitoids. Initially, parasitoids may use chemical information garnered from the microhabitat in which they emerge to locate hosts. Spalangia cameroni and Muscidifurax raptor are commercially available parasitoids of filth flies. Postemergence exposure to a specific manure may provide a way to increase parasitism in specific microhabitats found at livestock facilities upon release. In this study, female parasitoids of both species were exposed to equine manure, bovine manure, or clean pupae. Females from each emergence exposure were tested in a two-choice arena (house fly hosts in bovine manure versus clean pupae, equine manure versus clean pupae, and equine manure versus bovine manure) for progeny production. There was a detectable but small effect of postemergence exposure on S. cameroni, but it was not sufficient to reverse innate preferences. Females consistently produced more progeny in hosts found in any manure over clean pupae, and in equine manure over bovine manure. The effect of postemergence exposure on M. raptor was also detectable but small. Females produced equal numbers of progeny in bovine manure versus clean pupae, as opposed to preferring to oviposit in clean pupae as with all other treatments. Preferences by M. raptor were overall less marked than for S. cameroni; indeed most of the variability observed for this species did not result from the treatment design. Residual host mortality was also detectably altered by exposure in both species, but the effect was small. Thus, postemergence exposure did not consistently and effectively manipulate these parasitoids to producing progeny in different exposure manures, suggesting that microhabitat preferences are largely determined by other factors.

The objective of this study was to evaluate the suitability of killed house fly (Musca domestica L) pupae for production of two economically important pupal parasitoids. Two-day-old fly pupae were subjected to heat shock treatments of varying temperatures and durations in an oven at >or=70% RH; exposure to temperatures of 55 degrees C or higher for 15 min or longer resulted in 100% mortality. Exposure to 50 degrees C resulted in 40 and 91% mortality at 15 and 60 min, respectively. All (100%) pupae placed in a -80 degrees C freezer were killed after 10-min exposure; exposure times of

House flies are a ubiquitous insect that have the potential to spread many diseases to humans and livestock. Managing house fly populations is accomplished by having desirable baits, traps, and killing agents. Most house fly baits are designed for outdoor use or limited indoor use, and have a foul odor that is not conducive to food preparatory and dining areas. Blackstrap molasses has long been used as a house fly bait, but it is sticky and viscous, making it difficult to handle. This study sought to identify compounds present in blackstrap molasses that might be attractive to house flies, and therefore, provide the public with an indoor bait that does not have an offensive smell and is easy to handle. Indoor bioassays with house flies using 50% blackstrap molasses diluted in deionized water, a hexane extract of blackstrap molasses, and deionized water, elicited 86.2%, 70.6%, and 13.8% responses, respectively. Hexane and diethyl ether extracts of blackstrap molasses produced a large number of compounds with widely differing organic structures including substituted phenols, nitrogen and oxygen heterocycles, carboxylic acids, and many other organic compounds.

Filth flies are pests of livestock, and can transmit pathogens that cause disease to animals and their caretakers. Studies have shown successful infection of adult filth flies following exposure to different strains and formulations of entomopathogenic fungi. This study aimed to examine the effects of commercial formulations of Beauveria bassiana (Balsamo) (Moniliales: Moniliaceae) (i.e., BotaniGard ES, Mycotrol O, balEnce), and Metarhizium brunneum (Metsch.) (Ascomycota: Hypocreales) (i.e., Met52 EC), on filth fly oviposition and immature fly survival after exposure. House flies, Musca domestica L., laid significantly fewer eggs on Met52 EC-treated surfaces than on surfaces treated with all other products and the control. Similar numbers of eggs were laid on surfaces treated with all B. bassiana products, but egg production was half of the control. Stable flies, Stomoxys calcitrans (L.), laid the fewest eggs on Met52 EC- and Mycotrol O-treated surfaces. This species did not distinguish between the remaining products and the control. In a second experiment, house fly eggs were placed on treated cloths so that hatched larvae contacted the treatment prior to development. Met52 EC had the greatest effect on immature survival with a significant reduction in recovered pupae at the medium and high doses of fungi. Overall, Met52 EC, containing M. brunneum, had the greatest effect on house fly and stable fly oviposition deterrence and immature development of house flies. Management implications are discussed.

Release of parasitic wasps (Hymenoptera: Pteromalidae) as biological control agents for house flies and stable flies in livestock confinements has had variable success. In part, this may reflect a lack of knowledge regarding the optimal distance to be used between parasitoid release stations. In the current study, we assessed the effect of linear distance on host parasitism by the wasp Spalangia cameroni Perkins. In open fields at distances ranging from 1 m to 60 m from a central point, house fly puparia were placed in a mixture of pine shavings soiled with equine manure, urine, and alfalfa hay. Releases of S. cameroni then were made using a 5:1 host: parasitoid ratio. Host pupae were parasitized at all distances, with the highest rate of total parasitism (68.9%) recorded ≤ 5 m from the release site. Analyses of results using non-linear and linear models suggest that S. cameroni should be released in close proximity to host development areas. Additionally, releases may not be suitable in pasture situations where long-distance flight is required for control. However, further testing is needed to examine the effect of density-dependent dispersal and diffusion of S. cameroni.

Attack rates, progeny production, sex ratios, and host use efficiency of Muscidufurax raptor Girualt and Sanders, Spalangia cameroni Perkins, S. endius (Walker), S. nigroaenea Curtis, S. gemina Boucek (Hymenoptera: Pteromalidae), and Dirhinus himalayanus (Hymenoptera: Chalcididae) were evaluated in laboratory bioassays with five dipteran hosts: house fly (Musca domestica L.), stable fly (Stomoxys calcitrans L.), horn fly (Hematobia irritans L.), black dump fly [Hydrotaea aenescens (Weidemann)] (Diptera: Muscidae), and a flesh fly (Sarcophaga bullata Parker) (Diptera: Sarcophagidae). M. raptor, S.cameroni, and S. endius readily attacked and produced progeny on all five host species, with substantially lower production from S. bullata than from the muscid hosts. Rates of host attacks by S. nigroaenea and S. gemina were similar on house fly, stable fly, and black dump fly hosts, with lower rates on horn fly; almost no progeny were produced by S. nigroaenea on S. bullata hosts. D. himalayanus, a larg...

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 population...

The effects of an undetermined species of Nosema on fitness of the muscoid fly parasitoid Tachinaephagus zealandicus were examined in the laboratory. Infected female parasitoids that were given honey and water lived about one-half as long as uninfected parasitoids under these feeding conditions. Effects of infection on longevity were strongest at 30°C; infected and uninfected females lived 2.8 and 8.7 d, respectively. Infected and uninfected parasitoids that were given only water had similar longevities, but water-only–fed parasitoids had much shorter lifespans than honey-fed parasitoids at all temperatures. Infection did not result in significant lengthening of development times of immature stages, with male and female parasitoids completing development from egg to adult in ≈23, 33, and 60 d at 25, 20, and 15°C, respectively. Overall emergence of uninfected parasitoid adults was 16 times greater than infected parasitoids at 15°C. Emergence of uninfected parasitoids was 11 and 3 times greater tha...

Several methods were investigated for managing Nosema disease in the parasitoid Muscidifurax raptor Girault and Sanders. Treatment of parasitoid eggs or pupae within host puparia with gamma radiation from a cesium-137 source were either lethal to the parasitoids at all dosages tested (eggs) or failed to reduce infection rates (pupae). Exposure of parasitoid eggs within host puparia to heat was effective at several temperatures and exposure times. Optimal results for disease reduction were achieved with a 5-h exposure to 45°C, which resulted in no infection in the resulting adult parasitoids. Continuous rearing at elevated temperatures (30 and 32°C) for three generations resulted in decreased spore loads in infected parasitoids but did not reduce infection rates. Incorporation of the drugs albendazole and rifampicin into rearing media of the parasitoid’s host (house fly immatures) resulted in pupae that were of poor quality and did not reduce infection rates in parasitoids that developed in flies reared on drug-treated media. Treatment of adult parasitoids with 3% albendazole and/or rifampicin resulted in decreased rates of transovarial transmission of the disease. Transmission blockage required 3–7 d of exposure to the drug before substantial treatment effects were manifest. Parasitoids that fed for 7 d on rifampicin-treated honey transmitted the disease to 57.7% of their progeny compared with a 99.1% transmission rate among untreated parasitoids. An uninfected colony of M. raptor was established by pooling cured parasitoids from heat shock and drug treatment experiments. Parasitoids from the uninfected colony lived longer and produced over twice as many female progeny (201.2) as infected parasitoids (85.2).

Tachinaephagus zealandicus As hmead isa gregariousendoparas itoid that attacksthird instars of muscoid sies, including house sies, Musca domestica L. A colony of thisparas itoid was established from samples collected from a poultry farm in Santa Cruz da Conceicao, Sao Paulo, Brazil. The objective of this study was to evaluate the insuence of feeding treatment, host density and temperature on attack rateson T. zealandicus. Paras itoidsthat were given honey asadultsattacked two to three times as many house sy larvae (25 host attacks/female/d) as parasitoids that were given only water or nothing. Host attacks and progeny production by T. zealandicus on house sy and Chrysomyia putoria increased over the range of host:parasitoid ratios tested, reaching a maximum of 21Ð22 hosts killed and 13 progeny produced/female/d at the highest host density of 32 larvae/female. Host attacks were higher at 22C than at the other temperaturesstudied (20 Ð29 C), but differences in attack rateswere small over the range of 20 Ð27 C (10 Ð13 host attacks/female). Comparatively few hosts (6.3) were attacked at 29C. Higher rates of progeny production also were observed among parasitoids tested at lower temperatures (9 Ð11 progeny produced/female at 20 Ð22C) than at 29C (1.8 progeny/female). Femalesof T. zealandicus that were stored at 15C after emergence had highest rates of host attacks (58 Ð 62 hosts killed per group of Þve female parasitoids) and progeny production (174 Ð261 progeny) after 6 Ð12 d of storage at this temperature; relatively few hosts were attacked or parasitized (6 Ð9 host attacks and progeny/group) after 0 o r1da t 15C.

Tachinaephagus zealandicus Ashmead is a gregarious endoparasitoid that attacks third instars of muscoid flies in the Southern Hemisphere. The purpose of the current study was to evaluate the influence of six constant temperatures (16, 18, 20, 22, 25, and 27°C) on development time, the influence of emergence order on longevity, and the effects of temperature and food treatment on longevity. Emergence success was greatest at 22°C for both males and females; significantly fewer (24.1–30.4%) parasitoids emerged at 16 and 25°C compared with 22°C. Development time ranged from 24.0 to 56.9 d for both sexes. No emergence was observed at 27°C. Early-emerging parasitoids had greater longevity than parasitoids that emerged later from the same cohorts. The longevity of females given honey and water decreased with increasing temperature, and those reared at 16°C lived about three times longer than those kept at 27°C. Females given honey and water had similar longevities at 16–20°C, and females that were given only water lived for only 4.8–7.6 d at all temperatures. Females lived significantly longer overall than males at all temperatures except 16°C, but differences due to sex were small compared with the effects of temperature and nutrition. Further investigations will be necessary to determine the climatic zones in which T. zealandicus is most likely to be an effective biological control agent of muscoid flies.

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 ...