Production of Filth Fly Parasitoids (Hymenoptera: Pteromalidae) on Fresh and on Freeze-killed and Stored House Fly Pupae

Laboratory experiments were performed to assess the effects of age, status (fresh versus freezekilled), and storage regime on the suitability of house fly, Musca domestica L. (Diptera: Muscidae) pupae as hosts for Muscidifurax raptor Girault & Saunders, M. raptorellus Kogan & Legner, M. zaraptor Kogan & Legner, Spalangia cameroni Perkins, Trichomalopsis sarcophagae (Gahan) and Urolepis rufipes (Ashmead) (Hymenoptera: Pteromalidae). Production of all species was maximized on pupae aged 24 + h post-pupation. Fresh pupae could not be refrigerated at 10°C or less, or at 15°C without a significant decline in their suitability as hosts. Although production of S. cameroni was essentially limited to the use of fresh house fly pupae, M. raptor , M. raptorellus , M. zaraptor , T. sarcophagae and U. rufipes could be reared on either fresh or freeze-killed pupae stored at - 20 °C for up to 6 months prior to parasitism. The suitability of freeze-killed pupae declined during storage when used for production of male and female M. raptorellus and M. zaraptor , and possibly for male T. sarcophagae . No other effects of storage on parasitoid production were detected. These results suggest that insectaries can stockpile fly pupae in freezers during times of overproduction for future use in mass-rearing M. raptor, M. raptorellus, M. zaraptor, T. sarcophagae and U. rufipes as biocontrol agents of filth flies.     

Winter survival of nuisance fly parasitoids (Hymenoptera: Pteromalidae) in Canada and Denmark

Independent studies were performed in Canada and in Denmark to assess the survival of parasitic wasps (Hymenoptera: Pteromalidae) wintering in puparia of house fly, Musca domestica Linnaeus (Diptera: Muscidae). Data in Canada were collected for Muscidifurax raptorGirault & Saunders, M. raptorellus Kogan & Legner, M. zaraptor Kogan & Legner, Nasonia vitripennis(Walker), Spalangia cameroni Perkins, Trichomalopsis sarcophagae (Gahan) and Urolepis rufipes (Ashmead) in three microsites at an outdoor cattle facility in southern Alberta. Survival was highest for N. vitripennis, T. sarcophagae and U. rufipes, ranging from near zero to c. 7%. No survival was observed for S. cameroni. Daily mean values for ambient air temperature (DMAT) averaged about -3.5 degrees C during exposure periods. Data for Denmark were collected for M. raptor, S. cameroni and U. rufipes in a dairy barn and in a swine barn. Survival of M. raptor and U. rufipes was higher than that of S. cameroni in the dairy barn (DMAT = 8.6 degrees C), with the three species having similar survival in the swine barn (DMAT = 15.4 degrees C). In both studies, parasitoids in egg stages were least likely to survive. These results identify the potential for T. sarcophagae and U. rufipes to be commercialized for use in northern climates as biocontrol agents for nuisance flies, compare directly the cold-hardiness of commercialized species (i.e. all of the above species excluding T. sarcophagae and U. rufipes), and document the importance of microsite on winter survival.     

Commercial and naturally occurring fly parasitoids (Hymenoptera: Pteromalidae) as biological control agents of stable flies and house flies (Diptera: Muscidae) on California dairies

Filth fly parasites reared by commercial insectaries were released on two dairies (MO, DG) in southern California to determine their effect on populations of house flies, Musca domestica L., and stable flies, Stomoxys calcitrans (L.). Spalangia endius Walker, Muscidifurax raptorellus Kogan and Legner, and Muscidifurax zaraptor Kogan and Legner were released on the MO dairy from 1985 to 1987 in varying quantities. Parasitism by Muscidifurax zaraptor on the MO dairy was significantly higher (P less than 0.05) from the field-collected stable fly (4.4%) and house fly (12.5%) pupae, compared with a control dairy (0.1%, stable fly; 1.3%, house fly). Muscidifurax zaraptor, released from April through October during 1987 on the DG dairy (350,000 per month), was not recovered in a significantly higher proportion from either fly species relative to the corresponding control dairy. No specimens of Muscidifurax raptorellus were recovered from the MO dairy. Parasite treatments had no apparent effect on adult populations of either fly species or on overall parasitism rate of field-collected stable fly (16.8%, MO; 17.2%, DG) and house fly (23.3%, MO; 20.9%, DG) pupae. Spalangia spp. were the predominant parasites recovered from field-collected stable fly and house fly pupae on all four dairies. Sentinel house fly pupae placed in fly-breeding sites on both release dairies were parasitized at a significantly higher rate, as compared with sentinel pupae on control dairies. The generic composition of parasites emerging from sentinel house fly pupae was 20.6% Spalangia spp. and 73.2% Muscidifurax spp., whereas in field-collected house fly pupae, Spalangia spp. and Muscidifurax spp. constituted 74.3 and 19.6% of the parasites, respectively.     

Impact of exposure length and pupal source on Muscidifurax raptorellus and Nasonia vitripennis (Hymenoptera: Pteromalidae) parasitism in a New York poultry facility

Commercially obtained Nasonia vitripennis Walker and Muscidifurax raptorellus Kogan & Legner were released weekly for 12 wk into a high-rise, caged-layer poultry house. After the release period, parasitoids were sampled using sentinel house fly (Musca domestica L.) pupae that were either laboratory-reared or field-collected as larvae and exposed for 2, 4, 7, and 14 d. Parasitoid-induced mortality was observed in 31% of laboratory colony pupae and in 26% of field-collected pupae, whereas successful parasitism rates of 48 and 51% were observed from these pupal sources, respectively. Parasitism was primarily by M. raptorellus (88%), and Muscidifurax raptor Girault & Sanders (11%), while N. vitripennis accounted for <1%. Percent female progeny ranged from 43%, in M. raptorellus to 76% in N. vitripennis. Parasitoid emergence from 2-d exposed sentinel pupae was the lowest of all treatments. Parasitoid emergence from 7-d exposed sentinel pupae was the highest of all treatments. We found no differences between pupal source, suggesting that when sampling for M. raptor, M. raptorellus, and N. vitripennis, in poultry facilities, pupal source is not a confounding factor.     

Temperature-dependent development and parasitism rates of four species of Pteromalidae (Hymenoptera) parasitoids of house fly (Musca domestica) pupae

Parasitoid development, parasitoid-induced host mortality and parasitoid progeny emergence were determined at five constant temperatures for Muscidifurax raptor Girault and Sanders, Muscidifurax zaraptor Kogan and Legner, Spalangia cameroni Perkins and Spalangia endius Walker using pupae of the house fly, Musca domestica L., as hosts. At temperatures of 20, 25, 30 and 35 degrees C the median development times (days from oviposition to adult emergence), respectively, were M. raptor (28.4, 20.7, 14.3, 14.5), M. zaraptor (30.6, 22.8, 14.1, 14.2), S. cameroni (55.6, 35.2, 21.8, 25.0) and S. endius (52.4, 31.5, 16.3, 14.6). All species failed to emerge at 15 degrees C. Using densities of five parasitoids and 100 hosts and a 24 h exposure period, Muscidifurax species oviposited at a greater rate over a wider range of temperatures than Spalangia species. At 15, 20, 25, 30 and 35 degrees C the mean number of pupae killed per parasitoid were, respectively, M. raptor (1.4, 7.4, 10.5, 13.7, 14.1), M. zaraptor (0.0, 3.3, 8.9, 14.4, 15.0), S.cameroni (0.0, 7.8, 11.0, 11.9, 7.4), S.endius (0.6, 4.0, 7.5, 12.0, 11.7), and means of the number of parasitoid progeny per parasitoid were, respectively, M.raptor (0.2, 5.2, 7.9, 11.8, 11.6), M.zaraptor (1.3, 4.4, 8.2, 13.0, 13.7), S.cameroni (0.0, 2.4, 4.7, 5.1, 1.0), S.endius (0.0, 0.9, 3.4, 7.5, 4.9). Development and ovipositional activity in S.cameroni was strongly inhibited at 35 degrees C. The model by Sharpe & DeMichele (1977) was used to describe temperature-dependent development and the number of parasitoid progeny produced per parasitoid at temperatures of 15-30 degrees C in all species.     

Parasitism of house fly (Musca domestica) pupae by four species of Pteromalidae (Hymenoptera): effects of host–parasitoid densities and host distribution

Parasitoid-induced mortality of house fly, Musca domestica L., pupae and parasitoid progeny emergence by four species of pteromalid parasitoids, Muscidifurax raptor Girault & Sanders, M.zaraptor Kogan & Legner, Spalangia cameroni Perkins and S.endius Walker, were determined for a 24 h exposure period using parasitoid: host ratios ranging from 1:2 to 1:50. When the number of parasitoids was held constant (n = 5) and the numbers of hosts varied, and when the number of hosts was held constant (n = 100) and the number of parasitoids varied, both the number of pupae killed per parasitoid and the number of parasitoid progeny per parasitoid increased with increasing parasitoid:host ratios to reach an upper limit asymptotically. Maximum values were, respectively: M.raptor (14.7, 11.1), M.zaraptor (12.3, 9.3), S.cameroni (16.9, 5.5), S.endius (14.8, 9.7) with no consistent effects attributed to parasitoid interference. For M.raptor and S.cameroni at parasitoid:host ratios of 1:10, the pupal mortality and progeny emergence were determined for a 24 h exposure period when hosts were distributed in poultry manure at four levels of aggregation ranging from clumped to uniform. Pupal mortality was least in clumped distributions, while parasitoid progeny emergence was not significantly different.     

Evaluation of field propagation of Muscidifurax zaraptor (Hymenoptera: Pteromalidae) for control of flies associated with confined beef cattle.

The parasitic wasp Muscidifurax zaraptor Kogan & Legner was mass-reared in the field to control house flies, Musca domestica L., on two Nebraska beef cattle confinements. About 50,000 freeze-killed house fly pupae were exposed to a single release of M. zaraptor in the field. Placement of six additional cohorts of 50,000 freeze-killed pupae at the release sites at 2-wk intervals resulted in a mean parasite emergence of 56.4% over the study period. Mean fly mortality of 37.3 and 25.9% occurred in sentinel pupae placed around the perimeter of two release sites, compared with 3.9% for two control sites. We demonstrated a negative correlation between host reduction in sentinel cohorts and distances the cohorts were placed from parasite release sites. However, data indicated that other environmental factors also influenced the success of M. zaraptor in locating sentinel hosts. Correlation between mortality in sentinel pupae and numbers of parasites released was not evident. Temperatures above approximately 28 degrees C appeared to reduce the effectiveness of M. zaraptor.     

Early season dispersal of Muscidifurax zaraptor (Hymenoptera: Pteromalidae) utilizing freeze-killed housefly pupae as hosts

The pteromalid wasp, Muscidifurax zaraptor Kogan and Legner, was released at three locations at a dairy in May before housefly and stable fly breeding had begun. Freeze-killed housefly pupae were placed adjacent to the emerging parasites at biweekly intervals for a 6-week period. Hosts placed out weeks 0 and 2 were heavily parasitized. Decreased parasitism in hosts placed out at week 4 suggested that many of the M. zaraptor had dispersed or died. High parasitism of hosts placed in the field at week 6 was the result of second generation parasites emerging from pupae placed out at week 0. Parasitism of freeze-killed housefly pupae placed 6 m and in the four cardinal directions from the release points was similar but lower than for hosts placed adjacent to the emerging parasites. The study demonstrated that emerging M. zaraptor readily utilized nearby freeze-killed housefly pupae but the availability of these hosts did not deter the parasites from searching for additional hosts.     

The ability of selected pupal parasitoids (Hymenoptera: Pteromalidae) to locate stable fly hosts in a soiled equine bedding substrate

The ability of Spalangia cameroni Perkins, Spalangia endius Walker, and Muscidifurax raptorellus Kogan and Legner to locate and attack stable fly hosts was evaluated under laboratory conditions. Postfeeding third-instar stable fly larvae were released and allowed to pupate in two arena types: large 4.8 liter chambers containing a field-collected, soiled equine bedding substrate; or 120-ml plastic cups containing wood chips. At the time of fly pupariation, parasitoids were released and permitted 72 h to locate and attack hosts. On average, parasitism rates of freely accessible stable fly pupae in cups were not significantly different between parasitoid species. However, parasitism rates in chambers containing either Spalangia spp. were ≈50-fold more than M. raptorellus. Additional intraspecies analysis revealed that parasitism rates both by S. cameroni and S. endius were not significantly different when pupae were freely accessible or within bedding, whereas M. raptorellus attacked significantly more pupae in cups than in the larger chambers where hosts were distributed within bedding. These results suggest that Spalangia spp. are more suited to successfully locate and attack hosts in habitats created by equine husbandry in Florida. Therefore, commercially available parasitoid mixtures containing Muscidifurax spp. may be ineffective if used as a control measure at Florida equine facilities.     

Parasitism rates of Muscidifurax raptorellus and Nasonia vitripennis (Hymenoptera: Pteromalidae) after individual and paired releases in New York poultry facilities

Commercially reared parasitoids were released into three high-rise, caged-layer poultry houses; one house received only N. vitripennis Walker, the second house received only M. raptorellus Kogan & Legner, and the third house received an equal ratio of both species. Overall, house fly parasitism by M. raptorellus was never higher than 7% in any house. Most parasitism in the M. raptorellus release house was attributed to N. vitripennis. Parasitism of house fly pupae by M. raptorellus did not significantly increase during or after the 6-wk release period in the house that received both parasitoids. However, a depression in total parasitism was not detected when releases of the two species were made in this house.     

Influence of habitat and temperature on dispersal behaviour of two pteromalid parasitoids of houseflies during an inundative release at a dairy barn

1. About 11,000 each of Muscidifurax raptor Girault and Saunders and Urolepis rufipes (Ashmead) were released weekly for 7 weeks at a commercial dairy farm in central New York state, U.S.A. Dispersal behaviour was monitored by parasitism rates of house fly, Musca domestica L., pupae placed in sentinel bags. 2. M. raptor, which was released inside the barn, parasitized fly pupae both inside and outside, and it achieved highest rates of parasitism in indoor straw calf-bedding and in outdoor manure and silage. 3. U. rufipes, which was released outside the barn, did not attack pupae inside the barn, and its highest rates of parasitism occurred in outdoor manure and silage. 4. M. raptor appeared to be more effective than U. rufipes in parasitizing pupae located at sites where natural fly-breeding occurred. 5. Interspecific competition did not appear to explain these distribution patterns. 6. There was no significant trend in parasitism by M. raptor as a function of distance from the release station. Furthermore, high rates of parasitism near open doorways and at an outdoor site 30 m away suggests that M. raptor dispersed throughout the barn and its immediate surroundings. 7. Air temperature was positively correlated to flight activity, but not to parasitization activity in natural fly-breeding substrates.     

Recombinant males in the parasitic waspMuscidifurax raptorellus [Hymenoptera: pteromalidae]

The identification of recombinant males inMuscidifurax raptorellus Kogan & Legner, secured from virgin hybrid ♀♀ formed by crossing cohorts from solitary and gregarious, populations, supports chromosomal inheritance of gregarious oviposition behavior. Examination of parasitization behavior in female progeny that had recombinant male fathers, indicated the, existence of recombinant ♂♂. Such ♂♂ also were able to elicit immediate phenotypic changes in their female mating partners, at intensities expected from their genetic character, as previously observed with original parental ♂♂ in this species. Progeny originating from F₁ mothers and recombinant fathers demonstrated the highest levels of heterosis, as measured by increased parasitization rates and numbers of eggs laid. Although extranuclear inheritance has not been eliminated, its influence on phenotypic changes in progeny seems minimal.      

Competition between the filth fly parasitoids Muscidifurax raptor and M. raptorellus (Hymenoptera: Pteromalidae)

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.     

Identification ofMuscidifuraxspp., Parasitoids of Muscoid Flies, by Composition Patterns of Cuticular Hydrocarbons

Gas chromatographic analysis of cuticular hydrocarbons ofMuscidifuraxspp. adult females revealed species-specific patterns of composition that allowed identification ofMuscidifurax raptorGirault and Sanders,Muscidifurax zaraptorKogan and Legner, andMuscidifurax raptorellusKogan and Legner. A total of 18 components, all C29–C37 alkanes and methylalkanes, accounted for over 90% of the total cuticular hydrocarbons for all three species.Muscidifurax zaraptorwas characterized by a high ratio (11.9) of 3-MeC₃₁:internal Me₂C₃₅'s, whereas this ratio was <3 for the other species.Muscidifurax raptorelluswas characterized by a low (<1) 3-MeC₃₁:3,7,15-Me₃C₃₇ratio compared with ratios of 3.1 and 6.3 for these components inM. raptorandM. zaraptor, respectively. Three populations ofM. raptorelluscould be distinguished from one another based on two other component ratios (5- and 7-MeC31:3MeC32, 5- and 7-MeC31:3,7- to 3,15-Me₂C₃₃) with either 100% (Nebraska population) or 90% (Chilean and Peruvian populations) certainty. Comparison ofM. raptorcolonies established from five different locations (Florida, France, Germany, Brazil, Hungary) indicated that the hydrocarbon pattern was highly conserved in this species. A dichotomous key to species based on ratios of cuticular hydrocarbon components unambiguously classified the 50 samples ofMuscidifuraxspp. used to construct the key, plus five additional samples from different geographic locations.     

Diploidy restoration in Wolbachia-infected Muscidifurax uniraptor (Hymenoptera: Pteromalidae)

Thelytokous reproduction, where females produce diploid female offspring without fertilization, can be found in many insects. In some Hymenoptera species, thelytoky is induced by Wolbachia, a group of cytoplasmically inherited bacteria. We compare and contrast early embryonic development in the thelytokous parthenogenetic species Muscidifurax uniraptor with the development of unfertilized eggs of the closely related arrhenotokous species, Muscidifurax raptorellus. In the Wolbachia-infected parasitic wasp M. uniraptor, meiosis and the first mitotic division occur normally. Diploidy restoration is achieved following the completion of the first mitosis. This pattern differs in the timing of diploidy restoration from previously described cases of Wolbachia-associated thelytoky. Results presented here suggest that different cytogenetic mechanisms of diploidy restoration may occur in different species with Wolbachia-induced thelytoky.     

Coexistence of predatoryMuscina stabulans andophyra aenescens [Dipt.: Muscidae] with dipterous prey in poultry manure

Significant relationships between the emergence of adult predatory species,Ophyra aenescens (Wiedemann) andMuscina stabulans (Fallen) and prey,Musca domestica L.,Fannia femoralis (Stein) andF. canicularis (L.) existed on 10 poultry ranches in southern California. Predatory species may function in natural control at intervals throughout the year, especially on predominant prey,F. femoralis andF. canicularis. Dipterous species, especiallyO. aenescens andF. femoralis, were only moderately influenced by the surface area of breeding habitat available. Accuracy in the measurement of population trends in Diptere may be enhanced by increasing the number of samples at any given ranch rather than the addition of study ranches. Parasitism, ranging from 22.6–37.8% of collected puparia, involved principally 3 species,Muscidifurax zaraptor Kogan & Legner,M. raptor Girault & Sanders andSpalangia cameroni Perkins.      

Phenotypic expressions of polygenes inMuscidifurax raptorellus [Hym.: Pteromalidae], a synanthropic fly parasitoid

The precise phenotypic measurements, percent multiple oviposition and the number of parasitoids developed per host, can be used to assess quantitative genetic variation governing multiple oviposition and development in the muscoid Diptera parasitoid,Muscidifurax raptorellus Kogan & Legner. Evidence for polygenic control was based on the significance of correlations between expected genomic content and behavioral expression. Data accumulated from 8 oviposition days seem sufficient to measure accurately polygenic expression in this species.      

Parasitoid-induced mortality of house fly pupae by Pteromalid wasps in the laboratory

House fly, Musca domestica L., pupae were exposed to six species of pteromalid parasitoids, Muscidifurax zaraptor Kogan and Legner, M. raptor Girault and Sanders, M. raptorellus Kogan and Legner, Pachycrepoideus vindemiae (Rondani), Spalangia nigroaenea Curtis, and Urolepis rufipes Ashmead. Exposures were made for 48 h at six parasitoid-to-host ratios to measure the effect of parasitoid density on parasitoid-induced mortality (PIM) of hosts (excluding mortality as measured by parasitoid emergence). PIM was evident at all parasitoid-to-host ratios for all six species. Fly eclosion declined with a corresponding increase in the parasitoid-to-host ratio; the reverse was generally true for PIM. Parasitoid emergence increased initially with a corresponding increase in the parasitoid-to-host ratio to a point (depending on the parasitoid species), but then declined. The three Muscidifurax spp. and P. vindemiae exhibited similar behavior and generally avoided previously stung hosts until ovipositional restraints broke down at the higher parasitoid-to-host ratios. S. nigroaenea and U. rufipes exhibited little ovipositional restraint, resulting in a high proportion of PIM of hosts. Understanding factors that influence PIM will provide better evaluations of field releases of parasitoids to control flies and will aid in the development of the most economic procedures for large scale rearing of pteromalid parasitoids.     

Identification of Muscidifurax Spp. by Polymerase Chain Reaction–Restriction Fragment Length Polymorphism

Polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP) analysis of the nuclear ribosomal ITS-1 region was used to differentiate Muscidifurax (Hymenoptera: Pteromalidae) species which are parasitoids of filth fly pupae. Three restriction enzymes, Dpn II, Mse I, and Taq I, produced restriction patterns which were diagnostic for the four species analyzed, M. raptor, M. raptorellus, M. uniraptor, and M. zaraptor. Seven other restriction enzymes were able to differentiate one or more of the species and can be used alone, or in combination with other enzymes, to verify identifications. No intraspecific variation was observed among the populations examined. The utility of the PCR-RFLP technique compared with other molecular and biochemical diagnostic procedures is discussed.     

Estimations of number of active loci,dominance and heritability in polygenic inheritance of gregarious behavior inMuscidifurax raptorellus [Hymenoptera: Pteromalidae]

Estimates of the minimum number of genetic loci governing expressions of gregarious oviposition inMuscidifurax raptorellus Kogan & Legner were enabled by the verification of recombinant males, which were secured from virgin hybrid females, formed by crossing cohorts from solitary and gregarious populations. Examinations of parasitization behavior in female progeny that had F₁ male fathers indicated the presence of hybrids among the males. Estimates of gene number, made on the basis of variances in P₁, F₁, F₂ and backcross progeny, and by observing behavior in second and third order backcrosses, ranged from two to 19, with most between two and five. However, backcrossing data suggested that at least eight loci were actively segregating for this characteristic. Semi-dominance of the solitary trait (D=0.63 to 0.84), and unequal gene effects probably caused these gene estimated to be lower than the actual number. Estimates of the coefficient of heritability, in the broad sense based on parental and F₁ and F₂ variances indicated that variability of gregarious behavior in the experimental environment was influenced >60% by genotypic factors, offspring-parent regression analyses gave estimates >38%.