Laboratory transmission and in vivo incubation of Entomophthora muscae (Entomophthorales: Entomophthoracae) in the onion fly,Delia antiqua (Diptera: Anthomyiidae)

A high-moisture infection chamber was used for the in vivo transmission of Entomophthora muscae within laboratory populations of the onion fly, Delia antiqua. This cadaver-to-fly transmission procedure provided an average experimental infection rate well above 95%. Laboratory infection and temperature-dependent incubation rates of E. muscae were further examined in adult populations of D. antiqua. The time from initial exposure until host death and pathogen sporulation was accurately predicted using a second-order function of the incubation temperature. A developmental base temperature of approximately 5°C was estimated, with 105 degree-days being the average number of heat units required between host infection and death. E. muscae transmission between D. antiqua and D. platura, two insect pests typically associated with Michigan onion production, was verified under laboratory conditions.     

Courtship behavior and discrimination between potential mates by maleDelia antiqua (Diptera: Anthomyiidae)

The repertoire of courtship behaviors of male onion maggots,Delia antiqua (Meigen), in a laboratory bioassay chamber, was analyzed by direct observation and by video recordings, in conjunction with a multichannel event recorder. Seven courtship behaviors were categorized: inspection from the substrate, aerial inspection, contact from the substrate, contact from the air, genital alignment, copulation, and male-male interaction. The frequency distribution of copulation bouts was best described by a Poisson distribution; peak mating activity occurred about 1 h into the bioassay. The duration in copulo, however, was extremely variable. On average, males spent 30 sin copulo (n=183); <30% of bouts were >50 s. The ability of males to discriminate between sexes, sexually immature and mature females, and between females ofD. antiqua and the cabbage maggot,Delia radicum (L.), was most pronounced in the elements of genital alignment and attempted copulation. The courtship and mating behavior inD. antiqua is consistent with a sequence that relies initially primarily on indiscriminate visual recognition of a potential mate, followed by species-and sex-specific semiochemical recognition upon contact.     

Identification of odorant-binding protein genes expressed in the antennae and the legs of the onion fly,Delia antiqua (Diptera: Anthomyiidae)

The onion fly, Delia antiqua (Meigen), is a pest specialized to the onion, Allium cepa L., and some other Allium plants. Host odorants play an important role in the attraction of D. antiqua adults and stimulation of oviposition in females. Odorant-binding proteins (OBPs) may serve as a first step in the perception of these chemical cues. In this study, to identify all OBP genes expressed in the chemosensory tissues in D. antiqua, RNA-seq analysis was carried out. In addition to the seven OBP genes previously identified, we found eight novel OBPs. Comparisons with Drosophila melanogaster Meigen OBP genes revealed that these 15 D. antiqua OBPs cover the structural variety observed in D. melanogaster OBPs, including Plus C and Minus C OBPs. These results suggest that a relatively large repertoire of chemosensory genes is maintained even in a specialist feeder.     

Onion maggot feeding and development on heterogeneous sections of the onion bulb

When given a choice of feeding sites in laboratory arenas, Delia antiqua (Meigen) larvae exhibited a 3: 1 bias for internal vs outer sections of onion bulbs and distributed themselves non-randomly among identical preferred onion sections. Larval clustering did not appear to be correlated with increased fitness in the laboratory; larval development was identical across the range of densities from 1 to 50 larvae per onion. Larvae feeding on preferred internal sections developed 14% faster, were 38% heavier upon pupation, and were two times more likely to survive to the adult stage than larvae feeding on outer sections. D. antiqua larval food preference was therefore positively correlated with increased fitness.     

Field monitoring of onion maggot (Delia antiqua) fly through improved trapping

Onion maggot (Delia antiqua) is an economically important pest of Allium crops in temperate regions throughout the world. Management of this pest is necessary to achieve economic returns and depends on insecticide regimes and cultural management. Current cultural management especially altering planting date, field location and crop rotation depend on monitoring. We evaluated the effect of shape, size, colour and chemical attractants on trap catch of field populations of adult D. antiqua flies in upstate New York. White, large diameter, spherical traps in conjunction with Delia Lure attractant performed the best in attracting and catching D. antiqua adults. These results suggest an improved means of attracting and capturing D. antiqua populations which could be useful in monitoring efforts and development of attract and kill strategies for pest control.     

Companion planting – do aromatic plants disrupt host‐plant finding by the cabbage root fly and the onion fly more effectively than non‐aromatic plants?

Brassica and Allium host‐plants were each surrounded by four non‐host plants to determine how background plants affected host‐plant finding by the cabbage root fly (Delia radicum L.) and the onion fly [Delia antiqua (Meig.)] (Diptera: Anthomyiidae), respectively. The 24 non‐host plants tested in field‐cage experiments included garden ‘bedding’ plants, weeds, aromatic plants, companion plants, and one vegetable plant. Of the 20 non‐host plants that disrupted host‐plant finding by the cabbage root fly, fewest eggs (18% of check total) were laid on host plants surrounded by the weed Chenopodium album L., and most (64% of check total) on those surrounded by the weed Fumaria officinalis L. Of the 15 plants that disrupted host‐plant finding in the preliminary tests involving the onion fly, the most disruptive (8% of check total) was a green‐leaved variant of the bedding plant Pelargonium × hortorum L.H. Bail and the least disruptive (57% of check total) was the aromatic plant Mentha piperita × citrata (Ehrh.) Briq. Plant cultivars of Dahlia variabilis (Willd.) Desf. and Pelargonium×hortorum, selected for their reddish foliage, were less disruptive than comparable cultivars with green foliage. The only surrounding plants that did not disrupt oviposition by the cabbage root fly were the low‐growing scrambling plant Sallopia convolvulus L., the grey‐foliage plant Cineraria maritima L., and two plants, Lobularia maritima (L.) Desv. and Lobelia erinus L. which, from their profuse covering of small flowers, appeared to be white and blue, respectively. The leaf on which the fly landed had a considerable effect on subsequent behaviour. Flies that landed on a host plant searched the leaf surface in an excited manner, whereas those that landed on a non‐host plant remained more or less motionless. Before taking off again, the flies stayed 2–5 times as long on the leaf of a non‐host plant as on the leaf of a host plant. Host‐plant finding was affected by the size (weight, leaf area, height) of the surrounding non‐host plants. ‘Companion plants’ and aromatic plants were no more disruptive to either species of fly than the other plants tested. Disruption by all plants resulted from their green leaves, and not from their odours and/or tastes.     

Dividing the pie: differential dung pat size utilization by sympatric Haematobia irritans and Musca autumnalis

Horn flies [Haematobia irritans (Diptera: Muscidae) (L.)] and face flies [Musca autumnalis (Diptera: Muscidae) De Geer] use the same larval resource, but their interactions are poorly studied. Dung pats (n = 350) were core sampled in the summers of 2012 and 2013 from irrigated pastures in Pomona, California, U.S.A. (34°03′N, 117°48′W) and held for face fly and horn fly emergence. Surface areas and estimated weights were recorded for each whole pat. Almost half (42.0%) of the pat cores yielded neither fly, 29.7% yielded horn flies only, 12.9% yielded face flies only and 15.4% yielded both flies. Of the fly‐positive pats, surface area and mass were larger for face fly‐occupied pats, whereas horn fly‐occupied pats were smaller. Pats shared by the two species were intermediate. Horn flies per positive core were unaffected by the absence/presence of face flies, but half as many face flies emerged when pats were co‐inhabited by horn flies. Face flies inhabited larger pats, which might better resist heating and drying, to which they are susceptible; horn flies inhabited a broad pat size range. Horn fly tolerance of lower dung moisture probably allows horn flies to colonize and survive in a wide range of pats in dry areas like southern California.     

Dietary constraints on sexual activity, mating success, and survivorship of male Delia antiqua

The effects of protein-deprivation on the sexual activity and reproductive fitness of male onion flies, Delia antiqua (Meigen) (Diptera: Anthomyiidae), were investigated under laboratory conditions. The percentage of males inseminating gravid females, the magnitude of ovipositional response, and the total numbers of eggs deposited in 1:1 or 1:10 male:female matchings over two days was unaffected by deprivation of dietary protein. The LT₅₀'s (median survival time) for solitary males provided proteinaceous, sucrose, or water diets were 38.0, 25.8, and 6.0 days, respectively. Yet independent of diet effects, males lost 50% of their wing tissue by fragmentation after 26 days, suggesting that wing condition is more important in determining male reproductive fitness than longevity. Male mating frequency in single pairings with previtellogenic females deprived of proteinaceous diet for ten days was similar to that of gravid, protein-fed females. In no-choice and choice mating bioassays at a 10:1 female:male ratio, however, males inseminated significantly fewer previtellogenic than gravid females over 24 h. Despite evidence for male autogeny, removal of exogenous protein resources in the Allium agroecosystem may have important effects on the reproductive competency and fecundity of D. antiqua.     

Pasture dragging fails to reliably suppress the emergence of horn flies (Haematobia irritans) and face flies (Musca autumnalis) from dung pats in a Mid-Atlantic North American climate

The beef industry endures major economic losses from a complex of flies that feed on bovine blood and mucus. For cattle on pasture, the most important of these pests are horn flies (Haematobia irritans [L.] [Diptera: Muscidae]) and face flies (Musca autumnalis [Diptera: Muscidae] De Geer). Pasture dragging to spread manure pats has been promoted as a management tactic for these species because their larvae inhabit bovine manure pats, but the efficacy of this practice has not been empirically validated. Spreading pats might promote fly mortality through desiccation or overheating, but these processes are weather-dependent and warrant testing in disparate climates. We evaluated pasture dragging effects while monitoring for weather interactions throughout nine experiment rounds in summers of 2018 and 2020 in Pennsylvania, USA. The manure spreading treatments increased pat surface area up to 300% but failed to significantly reduce emergence of horn flies and face flies as compared to controls. In contrast, precipitation and temperature were significant predictors in fly emergence models. Surprisingly, face fly emergence was significantly elevated in dragged pats twice in 2020. These data call for a reevaluation of pasture dragging as a management technique for horn flies and face flies across a range of climates.     

Mortality and knockdown effects of imidacloprid and methomyl in house fly (Musca domestica L., Diptera: Muscidae) populations

In this study, the knockdown and mortality effects of imidacloprid and methomyl were investigated. The residual surface applications were carried out to determine the knockdown effects (KDt₅₀ and KDt₉₅) and mortality (LD₅₀ and LD₉₅) induced by each insecticide. For mortality comparisons, the susceptible house fly (Musca domestica L., Diptera: Muscidae) of a WHO population and three natural field‐collected M. domestica populations from Turkey were used. In conclusion, it was found that the resistance to imidacloprid and methomyl was significantly higher in the field populations when compared to the susceptible population from WHO. The results showed that applicators and pest management decision‐makers should control and conduct an integrated pest management strategy by including biological agents to prevent the development of high levels of resistance in the field populations.     

Molecular detection of Streptococcus equi subspecies equi in face flies (Musca autumnalis) collected during a strangles outbreak on a Thoroughbred farm

The objective of this study was to detect Streptococcus equi subspecies equi (S. equi) (Lactobacillales: Streptococcaceae) using quantitative polymerase chain reaction (qPCR) in flies collected from a farm with a documented outbreak of strangles. A total of 1856 face flies [Musca autumnalis (Diptera: Muscidae)] were collected using conventional fly traps. The flies were processed for nucleic acid purification and tested for the presence of S. equi by qPCR. A total of 10/1856 flies (0.54%) tested qPCR-positive for S. equi. The results may implicate the presence of face flies as a risk factor for the transmission of S. equi and highlight the need to institute proper husbandry measures, biosecurity protocols and fly control in order to reduce the potential for infection in at-risk horses.     

Evaluation of resistance to permethrin,cypermethrin and deltamethrin in different populations of Musca domestica (L.), collected from the Iranian dairy cattle farms

The house fly, Musca domestica (Linnaeus) (Diptera: Muscidae), is a major pest for human and livestock health and is also resistant to different insecticides. Herein, six M. domestica populations were collected, five of them from industrial cattle farms and the Koohrang population from a remote area as a susceptible population. The resistance/susceptibility of populations to three pyrethroids was evaluated. High levels of permethrin resistance were observed in all field populations and the resistance ratios (RRs) were estimated to vary from 52- to 129-fold. Resistant populations also exhibited resistance to other pyrethroids (cypermethrin and deltamethrin), with RRs ranging between 45- and 180-fold. According to synergistic (piperonyl butoxide, diethyl maleate and triphenyl phosphate) and enzymatic assays, resistant populations exhibited multiple resistance phenotypes. Cytochrome P450 monooxygenases (P450s), glutathione S-transferases (GSTs), and carboxylesterases (CarEs) were found to be involved in pyrethroid resistance in Isfahan population, P450s and GSTs in Mobarake population and CarEs detoxified pyrethroids in Natanz and Alavijeh populations. As substitution of Leucine (CTT) with Phenylalanine (TTT) at position 1014 of the voltage sensitive sodium channel (VSSC) gene is the most common mutation conferring resistance to pyrethroids in M. domestica, we sequenced a partial fragment of IIS6 and L1014F mutation was detected in all resistant populations. The present study provides valuable information for early detection of pyrethroid resistance and developing resistance management strategies in the house fly populations.     

Estimation of the effects of buffalo fly (Haematobia irritans exigua) on the milk production of dairy cattle based on a meta-analysis of literature data

Published reports on the effect of buffalo fly Haematobia irritans exigua De Meijere (Diptera: Muscidae) and the closely related horn fly (H. irritans) were examined and analysed using non-linear weighted regression techniques in an attempt to establish the relationship between daily production loss (D), average number of parasites (n) and the average damage per parasite per day (d), and to provide estimates of expected losses in milk yield (MYD) and live-weight gain (LWG) in dairy cattle. A Mitscherlich three-parameter model was used to explain the relationship between the total loss of production attributable to buffalo flies and the average number of flies associated with cattle. This model was significant (P<0.01), with R2 = 20.2% and predicted a threshold number of flies (n = 30) below which no adverse effects would be noted. At a moderate level of infestation (n = 200) dMYD was 2.6 ml/fly/day and dLWG was 0.14 g/fly/day, resulting in estimated daily losses in milk yield (D(MYD)) and live-weight gain (D(LWG)) of 520 ml and 28 g, respectively.     

Sensory interaction during trap-finding by female onion flies: implications for ovipositional host-plant finding

Using various three-dimensional traps alone and in combination with the onion volatile, dipropyl-disulphide (Pr₂S₂), we found that visual behaviour of female onion flies, Delia antiqua (Meigen), varied with the visual context (background composition and trap spacing) in which traps were presented and the females' reproductive state (mated vs. unmated). Against a background of real onions, females alighted more frequently on spherical than cylindrical traps, and white than green spheres, but females responded equally to white and green cylinders. In an onion field, baiting traps with Pr₂S₂ significantly increased female response to white over green traps, but had no influence on their response to trap shape. When traps were spaced 10 m apart and against a background of bare soil, females orienting to Pr₂S₂ baits alighted more frequently on vertical than horizontal traps, but shape and spectral reflectance were insignificant. However, when traps were spaced 0.25 m apart, females orienting to Pr₂S₂ baits alighted more frequently on cylinders than spheres. Mated females alighted more frequently on green than white cylinders, but unmated females responded to cylinders independent of spectral reflectance. When located 20 m downwind from Pr₂S₂ baits, mated females alighted on green cylinders significantly more often than unmated females. Response to traps mimicking onion plants suggests that ovipositional host-finding in female onion flies is dominated by olfactory responses at long range (several metres) and by visual cues at short-range (ca. 1 m). The view that host location by female onion flies is a hierarchical response pattern mediated by multiple sensory modalities and modified by resource level (habitat) and reproductive status, is discussed.     

Blowfly succession from possum (Trichosurus vulpecula) carrion in a sheep-farming zone

The significance of brushtail possum, Trichosurus vulpecula Kerr (Diprotodontia: Phalangeridae) carcasses to the succession and production of Diptera species and its relevance to fly strike management in Tasmania, Australia was examined. Calliphora stygia (Fabricius), Lucilia sericata (Meigen) and Calliphora vicina Robineau-Desvoidy (Diptera: Calliphoridae) were found to be the most abundant and Lucilia cuprina (Wiedemann) (Diptera: Calliphoridae) always the least abundant (< 1%) of the putative primary fly invading species to emerge. Carcasses that were left for up to 15 days in the field before being exposed to flies for 2 days also acted as breeding sites for large numbers of all primary fly species, with the exception of L. cuprina. Ordination analysis revealed no relationship between possum carcasses according to their length of exposure but did show significant negative associations between the number of putative secondary invaders (Chrysomya rufifacies (Macquart) (Diptera: Calliphoridae), Chrysomya varipes (Macquart) (Diptera: Calliphoridae) and putative tertiary flies (Hydrotaea rostrata Robineau-Desvoidy (Diptera: Muscidae)) to the number of C. vicina or C. stygia to emerge. There was enormous variability in the numbers of secondary/tertiary fly species to emerge from carcasses (0-11 450) that negatively correlated with the proportion of all flies to emerge that were primary, and with the mean size of adult L. sericata. Although carcass temperatures, especially those with a large larval population, were elevated, this did not appear to result in significant pre-adult fly mortality. The most important primary fly strike species L. cuprina was only found in insignificant numbers, whereas three other members of the fly strike fauna C. stygia, L. sericata and Ch. rufifacies did use possum carrion as an important breeding resource, but left implications for fly strike management inconclusive.     

Larvicidal potential of the polyol sweeteners erythritol and xylitol in two filth fly species

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 2^nd 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 2^nd 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.     

Chaperonin Contributes to Cold Hardiness of the Onion Maggot Delia antiqua through Repression of Depolymerization of Actin at Low Temperatures

Winter-diapause and cold-acclimated non-diapause pupae of the onion maggot, Delia antiqua (Diptera: Anthomyiidae), show strong cold hardiness. To obtain insights into the mechanisms involved in the enhancement of cold hardiness, we investigated the expression patterns of genes encoding subunits of chaperonin (CCT) and the morphology of actin, a substrate of CCT, at low temperatures. Quantitative real-time PCR analyses showed the mRNA levels of CCT subunits in pupal tissues to be highly correlated with the cold hardiness of the pupae. While actin in the Malpighian tubules of non-cold-hardy pupae showed extensive depolymerization after a cold treatment, actin in the same tissue of cold-hardy pupae was not depolymerized. Damage to cell membranes became apparent after the depolymerization of actin. Moreover, administration of Latrunculin B, an inhibitor of actin polymerization, to the larvae markedly decreased the cold hardiness of the pupae obtained. These findings suggest that CCT contributes to the cold hardiness of D. antiqua through the repression of depolymerization of actin at low temperatures.     

Identity and bioactivity of oviposition deterrents in pine oil for the onion maggot, Delia antiqua

The oviposition deterrent properties of pine oil (Norpine 65, Northwest Petrochemicals, Anacortes, Washington) for the onion maggot, Delia antiqua (Meigen), were verified using a two-choice bioassay with onion oil as an attractive control. The principal deterrent property of this pine oil was found to reside in three monoterpenes, 3-carene, limonene, and p-cymene, which were the primary constituents identified in the most deterrent of two fractions made by preparative gas chromatography of steam-distilled pine oil. At release rates of 220, 320 and 320 g per 24 h in two-choice bioassays these monoterpenes respectively caused 73.2, 65.4 and 56.3% deterrency of oviposition, while the ternary mixture released at 320 g per 24 h caused 88.6% deterrency. The ternary mixture also caused 62.5% deterrency in a no-choice bioassay. Of eight other monoterpenes tested, myrcene, -phellandrene, -terpinene, -phellandrene, -terpinene, terpinolene, and -pinene were significantly deterrent in declining order, while -pinene was inactive. The ternary mixture was released from glass capillary tubes or flexible plastic cylinders in further bioassays that challenged caged females to oviposit around the base of 35 potted onion seedlings with release devices placed on the soil surface. The most effective deterrency (85.3%) was achieved at a release rate of 280 g per 24 h per pot if plastic cylinder devices were deployed 24 h before the treated pot was exposed to D. antiqua females. If female D. antiqua were given only a treated pot, deterrency of oviposition on potted onion seedlings was significant, but low (11.7–63.2%). Because of incomplete efficacy, a monoterpene-based deterrent formulation would be best used operationally if combined with other deterrents, or if it were integrated with some other tactic.     

Sensilla of the larvae of Delia radicum L., D. Floralis F. and D. Antiqua MG. (diptera : Anthomyiidae)

A comparative SEM study of the sensilla of larval Delia radicum, D. floralis and D. antiqua (Diptera : Anthomyiidae) was undertaken. All had a dorsal organ and an anterior organ. These may contain olfactory receptors, contact chemo- and mechanoreceptors. Delia radicum and D. floralis also possessed a ventral organ, which is a possible chemo- and mechanoreceptor. Campaniform (mechanoreceptor) and hair sensilla, which may be humidity receptors, were present on the body segments.     

Temperature cycle amplitude alters the adult eclosion time and expression pattern of the circadian clock gene period in the onion fly

Soil temperature cycles are considered to play an important role in the entrainment of circadian clocks of underground insects. However, because of the low conductivity of soil, temperature cycles are gradually dampened and the phase of the temperature cycle is delayed with increasing soil depth. The onion fly, Delia antiqua, pupates at various soil depths, and its eclosion is timed by a circadian clock. This fly is able to compensate for the depth-dependent phase delay of temperature change by advancing the eclosion time with decreasing amplitude of the temperature cycle. Therefore, pupae can eclose at the appropriate time irrespective of their location at any depth. However, the mechanism that regulates eclosion time in response to temperature amplitude is still unknown. To understand whether this mechanism involves the circadian clock or further downstream physiological processes, we examined the expression patterns of period (per), a circadian clock gene, of D. antiqua under temperature cycles that were square wave cycles of 12-h warm phase (W) and 12-h cool phase (C) with the temperature difference of 8 °C (WC 29:21 °C) and 1 °C (WC 25.5:24.5 °C). The phase of oscillation in per expression was found to commence 3.5 h earlier under WC 25.5:24.5 °C as compared to WC 29:21 °C. This difference was in close agreement with the eclosion time difference between the two temperature cycles, suggesting that the mechanism that responds to the temperature amplitude involves the circadian clock.