Spatial and temporal habitat partitioning by calliphorid blowflies

Calliphorid blowflies perform an essential ecosystem service in the consumption, recycling and dispersion of carrion nutrients and are considered amongst the most important functional groups in an ecosystem. Some species are of economic importance as facultative agents of livestock myiasis. The interspecific ecological differences that facilitate coexistence within the blowfly community are not fully understood. The aim of this work was to quantify differences in habitat use by calliphorid species. Thirty traps were distributed among three habitats at two sites in southwest England for collections made during March–August 2016. A total of 17 246 specimens were caught, of which 2427 were Lucilia sericata, 51 Lucilia richardsi, 6580 Lucilia caesar, 307 Lucilia ampullacea, 4881 Calliphora vicina and 2959 Calliphora vomitoria (all: Diptera: Calliphoridae). Lucilia sericata was the dominant species in open habitats, whereas L. caesar was the most abundant species in shaded habitats. Calliphora specimens were more abundant in the cooler months. These findings suggest that Calliphora and Lucilia species show strong temporal segregation mediated by temperature, and that species of the genus Lucilia show differences in the use of habitats that are likely to be driven by differences in humidity tolerance and light intensity. These factors in combination result in effective niche partitioning.     

Behavioural response of Lucilia sericata to a decaying body infested by necrophagous insects

Numerous insect species are necrophagous, with Dipterans and Coleopterans being the most abundant on a corpse. Whether the presence of necrophageous species on a corpse affects the attraction of adult blowflies to the corpse is sparsely studied. We test the hypothesis that Lucilia sericata can discriminate the odour of a noncolonized cadaver from the odour of a cadaver on which conspecific and/or heterospecific necrophagous insects are feeding. The volatile organic compounds are collected from decomposing rats under four modalities: (i) in the absence of insects; (ii) in the presence of L. sericata adults; (iii) in the presence of Dermestes frischii adults; (iv) and in the presence of both D. frischii adults and L. sericata adults. During a multiple‐choice bioassay, blowflies are exposed to the four odour samples, and are shown to prefer the odour of a corpse where conspecific larvae are present. We also expect blowflies to avoid cadavers on which predators where present, although L. sericata are not repulsed by the odour of a cadaver colonized by hide beetles. We then compare the average quantities of all 61 volatile molecules identified, finding that the presence of necrophagous insects impacts some of them. However, none of the volatile organic compounds previously reported in the literature as being attractive for L. sericata adults are impacted by the presence of necrophagous insects. The results of the present study suggest that blowfly larvae modify the volatilome of a corpse, enabling adults to discriminate a colonized from a noncolonized corpse.     

Efficient germ-line transformation of the economically important pest species Lucilia cuprina and Lucilia sericata (Diptera, Calliphoridae)

The green blowfly species Lucilia cuprina and Lucilia sericata are economically important pests for the sheep industries of Australia and New Zealand. L. cuprina has long been considered a good target for a genetic pest management program. In addition, L. sericata maggots are used in the cleaning of wounds and necrotic tissue of patients suffering from ulcers that are difficult to treat by other methods. Development of efficient transgenesis methods would greatly facilitate the development of strains ideal for genetic control programs or could potentially improve “maggot therapy”. We have previously reported the germ-line transformation of L. cuprina and the design of a “female killing system” that could potentially be applied to this species. However, the efficiency of transformation obtained was low and transformed lines were difficult to detect due to the low expression of the EGFP marker used. Here we describe an efficient and reliable method for germ-line transformation of L. cuprina using new piggyBac vector and helper plasmids containing the strong promoter from the L. cuprina hsp83 gene to drive expression of the transposase and fluorescent protein marker gene. We also report, for the first time, the germ-line transformation of L. sericata using the new piggyBac vector/helper combination.     

The effects on genetic variability following a recent colonization event: the Australian sheep blowfly, Lucilia cuprina arrives in New Zealand

The Australian sheep blowfly, Lucilia cuprina, was first identified in New Zealand in 1988 and is now found to have spread throughout many sheep-farming regions. L. cuprina is estimated to have been present in New Zealand < 20 years, while in Australia L. cuprina has been estimated present > 100 years. The aim of this study was to determine the genetic effects of colonization of L. cuprina and to compare populations of L. cuprina from these two countries in terms of genetic variability and differentiation. Allozyme electrophoresis was used which revealed variability at six loci. 1680 blowflies were examined from 56 sites throughout L. cuprina's range in both countries. Genetic variability at each locus in terms of allele composition was found to be high and genetic differentiation varied considerably in New Zealand in comparison to Australia. Temporal sampling in New Zealand suggests seasonal fluctuations of population size in the recently colonized region of the South Island.     

Quickie well done: no evidence of physiological costs in the development race of Lucilia sericata necrophagous larvae

The present study focuses on trade-offs between the development rates and their life-history traits of feeding larvae. Indeed, quick growth is considered to be vital for necrophagous insects such as larvae because they are part of a rapidly changing ecosystem and at the mercy of many predators. However, excessively quick growth can have a negative effect on other life-history traits (e.g. survivorship and body size). The blowfly Lucilia sericata (Diptera: Calliphoridae) is used in the present study because this species is frequently found on carcasses in Central Europe and is a well-known experimental model in insect physiology and ecology. Individuals are reared from first instars to adults at two different constant temperatures (i.e. 15 and 28 °C) and under three different conditions: 100 Lucilia sericata (i.e. small monospecific condition), 250 L. sericata (i.e. large monospecific) or 125 L. sericata + 125 Calliphora vicina (i.e. heterospecific). The development time, pupal and larval survival rate and pupal size are determined individually under each condition. Regarding size and development time, substantial variation is observed between the different growth conditions and within a larval group under the same conditions. However, no trade-offs between development rate and size or survival are detected. These results demonstrate that, under the range of developmental conditions tested in the present study, the quick development of L. sericata larvae does not affect their size or mortality. This developmental plasticity may explain the evolutionary success of this species, which is present in several ecosystems worldwide and dominates the fresh-carrion ecosystem.     

Comparison of traps for the control of sheep blowfly in the U.K.

The ability of three commercially available trap types to catch Lucilia (Diptera: Calliphoridae) blowflies was assessed on three sheep farms in southwest England in 2008. The aim was to evaluate their relative value for the control of ovine cutaneous myiasis (sheep blowfly strike) on farms. There was a highly significant difference between the total number of female Lucilia caught per day by the traps, with an Agrilure Trap (Agrimin Ltd, Brigg, U.K.) catching more than the other trap types (Rescue Disposable Fly Trap, Sterling International, Spokane, U.S.A.; Redtop Trap, Miller Methods, Johannesburg, South Africa). However, there was no significant difference between the traps in the numbers of female Lucilia sericata (Meigen) caught. Nevertheless, consideration of the rate at which female L. sericata were caught over time showed that the Agrilure trap did not begin catching until about 30 days after its initial deployment. It subsequently caught L. sericata at a faster rate than the other two traps. The data suggest that the freeze‐dried liver bait used in the Agrilure trap required a period of about 30 days to become fully rehydrated and decompose to the degree required to attract and catch L. sericata. Once the bait was attractive, however, the trap outperformed the other two traps in terms of the rate of L. sericata capture. The Agrilure trap would appear to be the most effective of the designs tested for use against sheep blowfly and blowfly strike in the U.K., but care would be needed to ensure that the traps were deployed in advance of the blowfly season so that the bait was suitably aged when trapping was required.     

Morphological identification of Lucilia sericata,Lucilia cuprina and their hybrids (Diptera,Calliphoridae)

Hybrids of Lucilia sericata and Lucilia cuprina have been shown to exist in previous studies using molecular methods, but no study has shown explicitly that these hybrids can be identified morphologically. Published morphological characters used to identify L. sericata and L. cuprina were reviewed, and then scored and tested using specimens of both species and known hybrids. Ordination by multi-dimensional scaling indicated that the species were separable, and that hybrids resembled L. cuprina, whatever their origin. Discriminant function analysis of the characters successfully separated the specimens into three unambiguous groups – L. sericata, L. cuprina and hybrids. The hybrids were morphologically similar irrespective of whether they were from an ancient introgressed lineage or more modern. This is the first evidence that hybrids of these two species can be identified from their morphology. The usefulness of the morphological characters is also discussed and photographs of several characters are included to facilitate their assessment.     

In vitro antibacterial activity and physicochemical properties of a crude methanol extract of the larvae of the blow fly Lucilia cuprina

The emergence of multidrug‐resistant bacterial strains has prompted the reintroduction of maggot therapy in the treatment of chronic, infected wounds. Many previous studies have demonstrated the potent antibacterial activity of larval excretions/secretions of the blowfly Lucilia sericata (Meigen) (Diptera:Calliphoridae) against bacteria. However, the antibacterial activity of its sibling species, Lucilia cuprina (Wiedemann) (Diptera:Calliphoridae) against a wide range of pathogenic bacteria has never been determined. The aim of this study was to develop a new procedure to produce whole body extract of larvae of L. cuprina via methanol extraction as well as to demonstrate the in vitro antibacterial activity of this extract against seven selected wound pathogens (Staphylococcus aureus, methicillin‐resistant S. aureus, S. epidermidis, Streptococcus pyogenes, Klebsiella pneumoniae, Pseudomonas aeruginosa and Escherichia coli). The turbidimetric assay demonstrated that L. cuprina larval extract was significantly potent against all bacteria tested (P < 0.001). Additionally, colony‐forming unit (CFU), agar well diffusion and minimum inhibitory concentration assays have confirmed the apparent potency of larval extract against P. aeruginosa. The reconstituted larval extract was highly robust and thermally stable. These observations substantiated the feasibility of the methanol extraction method in the production of larval extract.     

Development and evaluation of male-only strains of the Australian sheep blowfly, <Emphasis Type="Italic">Lucilia cuprina</Emphasis>

The Australian sheep blowfly Lucilia cuprina (Wiedemann) is a major pest of sheep in Australia and New Zealand. From the 1960s to the 1980s there was a major effort to develop "field female killing" or FFK strains of L. cuprina that could be used for a cost-effective genetic control program. The FFK strains carried eye color mutations that were lethal to females in the field but not under conditions in the mass rearing facility. Males did not die in the field as normal copies of the eye color genes had been translocated to the Y chromosome and an autosome. Although the FFK strains showed some promise in field tests, a genetic control program in mainland Australia was never implemented for several reasons including instability of the FFK strains during mass rearing. A stable transgenic strain of L. cuprina that carried one or more dominant repressible female lethal genes offered the potential for efficient genetic control of blowfly populations. Here I review our research on tetracycline-repressible female lethal genetic systems, Lucilia germ-line transformation and sex determination genes that ultimately led to the successful development of transgenic "male-only" strains of L. cuprina. The technology developed for L. cuprina should be directly transferable to other blowfly livestock pests including L. sericata and the New World and Old World screwworm. 29     

Asymmetric competition between larvae of the blowflies Calliphora vicina and Lucilia sericata in carrion

1. The relative abundance of the blowflies Calliphora vicina (R.-D.) and Lucilia sericata (Meigen) in carrion was considered in relation to inter- and intraspecific larval competition and the distribution of adults between habitat types. 2. In mixed and pure laboratory cultures of L. sericata and C. vicina the mortality of both species increased and adult size declined as the initial larval number was increased. However, for L. sericata at initial numbers greater than ten larvae/g of liver, the effects of competition on adult size and mortality were greater in the mixed cultures than in the pure cultures. In contrast, for C. vicina at numbers greater than ten larvae/g of liver, the effects of competition on adult size and mortality were greater in the pure cultures than in the mixed cultures. 3. The laboratory data suggest therefore that for L. sericata the effects of interspecific competition with C. vicina on size and mortality were greater than the effects of intraspecific competition in a pure culture at the same initial number. Notably, however, the intensity of interspecific competition was not sufficiently asymmetric to allow C. vicina to exclude L. sericata even at the highest numbers examined. 4. In the field, higher numbers of adult L. sericata emerged from the carcasses of laboratory mice placed in open pasture than in woodland or hedgerow sites. In contrast, higher numbers of C. vicina emerged from carcasses placed in woodland and hedgerow sites. 5. Trapping showed that in the field adult L. sericata were relatively more abundant in open pasture than in woodland and hedgerow sites, while C. vicina were more abundant in woodland and hedgerow sites than in open pasture. 6. It is concluded that the low numbers of L. sericata that emerge from carrion relative to the numbers of C. vicina may, in part, be the result of asymmetric interspecific competition, but that the uneven distribution of adults of the two species among habitat types also plays a major role in shaping the blowfly community in carrion.     

Pollination activity by blowflies and honeybees on onions in breeders' cages

Honeybees (Apis mellifera) and three species of blowflies (Calliphora vomitoria, Lucilia caesar and L. sericata) were observed on mass-pollinated populations of onions (Allium cepa) in 4·4 × 3·6 m × 2-2·5 m cages in June-August 1977. Pollination activity at temperatures from 14° to 28°C was compared on the basis of mean time per flower touched on each umbel visit. Honeybees did not forage below 16°C. Above 16°C their mean time per flower was short (1·4 s) and varied little with temperature. For blowflies, it decreased markedly from 12·1 s at 14–15·5°C to 2·7 s at 26°C and above, largely because at low temperatures flies spent long periods quiescent or grooming rather than actively feeding. When flowering was not completely synchronous between cultivars, honeybees were more selective than blowflies, but where flowering was synchronous, both types of pollinator visited the two cultivars at random. For the size of cage used, neither type of insect had a distinct advantage as a pollinator of onions, despite their different behaviour patterns.     

Saccadic head and thorax movements in freely walking blowflies

Visual information processing is adapted to the statistics of natural visual stimuli, and these statistics depend to a large extent on the movements of an animal itself. To investigate such movements in freely walking blowflies, we measured the orientation and position of their head and thorax, with high spatial and temporal accuracy. Experiments were performed on Calliphora vicina, Lucilia cuprina and L. caesar. We found that thorax and head orientation of walking flies is typically different from the direction of walking, with differences of 45° common. During walking, the head and the thorax turn abruptly, with a frequency of 5–10 Hz and angular velocities in the order of 1,000°/s. These saccades are stereotyped: head and thorax start simultaneously, with the head turning faster, and finishing its turn before the thorax. The changes in position during walking are saccade-like as well, occurring synchronously, but on average slightly after the orientation saccades. Between orientation saccades the angular velocities are low and the head is held more stable than the thorax. We argue that the strategy of turning by saccades improves the performance of the visual system of blowflies.     

Thermoregulation in gregarious dipteran larvae: evidence of species‐specific temperature selection

Due to the ephemeral nature of carcasses they grow on, necrophagous blowfly larvae should minimize the time spent on the cadaver. This could be achieved by moving to high‐temperature areas. On that basis, we theorized that larvae placed in a heterogeneous thermal environment would move to the higher temperature that speed up their development. This study was designed to (1) test the ability of necrophagous larvae to orientate in a heterogeneous thermal environment, and (2) compare the temperatures selected by the larvae of three common blowfly species: Lucilia sericata (Meigen), Calliphora vomitoria (L.) and Calliphora vicina (Robineau‐Desvoidy). For this purpose, we designed a setup we named Thermograde. It consists of a food‐supplied linear thermal gradient that allows larvae to move, feed, and grow in close‐to‐real conditions, and to choose to stay at a given temperature. For each species and replication, 80 young third instars were placed on the thermal gradient. The location of larvae was observed after 19 h, with fifteen replications per species. The larvae of each species formed aggregations that were always located at the same temperatures, which were highly species‐specific: 33.3 ± 1.52 °C for L. sericata, 29.6 ± 1.63 °C for C. vomitoria, and 22.4 ± 1.55 °C for C. vicina. According to the literature, these value allows a fast development of the larvae, but not to reach the maximum development rate. As control experiments clearly demonstrate that larval distribution was not due to differences in food quality, we hypothesized that the local temperature selection by larvae may result from a trade‐off between development quality and duration. Indeed, temperature controls not only the development rate of the larvae, but also the quality of their growth and survival rate. Finally, results raise questions regarding the way larvae moved on the gradient and located their preferential temperature.     

Seasonal and spatial changes in blowfly production from small and large carcasses at Durham in lowland northeast England

Colonization by blowflies (Diptera: Calliphoridae) of mouse carcasses exposed in open agricultural land near Durham (54 45'N) changed from early spring monopolization by Calliphora vicina R.-D. to a summer pattern of multiple species exploitation by this species together with Lucilia caesar L., L. illustris Mg., L. silvarum Mg., L. sericata Mg. and L. richardsi Collin. In a garden at the edge of Durham, mouse carcasses were dominated by C. vicina from spring to autumn. Difference in mouse colonization between the agricultural and garden sites seemed to reflect differences in the blowfly species present, as measured by baited trap catches at the sites. In sets of C. vicina reared from mice under conditions of competition for larval food, it was found that resulting females were significantly larger than males, size being measured as mean wing length. Blowfly production from three sheep carcasses exposed successively at the agricultural site was dominated by C. vomitoria L. and L. caesar, but also produced other Lucilia species in small numbers, including L. sericata. These L. sericata females from sheep that had died from causes other than myiasis included full-sized specimens, in contrast to those produced from mouse carcasses that were all undersized individuals. As L. sericata females trapped on sheep pastures are predominantly full-sized, this suggests that large carcasses may, in part, be a source of the L. sericata population that attacks sheep as a myiasis agent. The nature of large carcasses as possible sources of L. sericata in lowland Britain is discussed.     

A simple,heat‐sterilizable artificial diet excluding animal‐derived ingredients for adult blowfly,Lucilia sericata

Artificial diets have been developed for Lucilia sericata (Meigen) blowfly larvae; however, diets for adults have not yet been developed. An adult diet that excludes animal tissues and animal‐derived ingredients and promotes not only ovarian development, but also oviposition, would aid in basic research and maggot debridement therapy. We have successfully developed artificial diets that exclude animal tissues and animal‐derived ingredients for L. sericata adults. The outcomes of the diets were comparable with those of a beef liver diet in terms of oviposition, adult survival and number of offspring.     

Isolation and characterization of polymorphic microsatellite markers in the blowflies Lucilia illustris and Lucilia sericata

Thirteen polymorphic microsatellite loci were developed for blowflies for use in studies of genetic differentiation in wild populations of Lucilia illustris, to detect the possible occurrence of bottlenecks and to study changes in genetic variation in laboratory populations of Lucilia sericata following artificial bottlenecks. In this preliminary study it was revealed that heterozygosity was lower than expected in wild populations and genetic variation had been lost in the laboratory population despite being kept at a large size.     

A new method for the production of sterile colonies of Lucilia sericata

Maggot debridement therapy (MDT) refers to the use of blowfly larvae to clean or debride an infected wound. Most commonly, larvae of Lucilia sericata (Meigen) (Diptera: Calliphoridae) are used, and are sterilized prior to use to ensure no further bacterial infections are introduced during treatment. Current methods sterilize eggs from laboratory‐reared blowfly colonies, after which sterile early second instar maggots can be provided to hospitals for use in treatment. Maggots not required for treatment are used for colony regeneration, in which sterility is not maintained. The ability to maintain sterility beyond this would allow further research into fly–bacteria interactions and the effects of different bacteria on the blowfly lifecycle. This study aimed to produce a colony of sterile adults, using current egg sterilization practice, but maintaining sterility through to pupation and emergence. The production of a sterile colony allows further research into the impact of bacteria on fly development and survival. Eggs were placed on a sterile food source within autoclaved plant tissue culture containers to allow growth under sterile conditions. Nutrient agar plating of sterilized and non‐sterilized eggs, larvae and adults (post‐emergence), as well as the pupation medium and feed source in nutrient broth confirmed the aerobic sterility of all samples involved. The lifecycle of L. sericata was successfully completed through pupation to emergence with no effects on lifespan or oviposition by the newly emerged, sterile adult colony.     

Contrasting responses to object orientation and illumination by the blowflies Chrysomya chloropyga and Lucilia sericata

The alighting response of two species of blowfly, Chrysomya chloropyga (Weidemann) and Lucilia sericata(Meigen) (Diptera: Calliphoridae), were considered in relation to the orientation and illumination of a flat, oblong visual object. A strong response to orientation was displayed by L. sericata with the greatest number alighting on the vertical object; there was no significant effect of orientation on the number of C. chloropyga which alighted. Both species alighted preferentially on the more strongly illuminated side of a vertical object. With the object horizontal and more strongly illuminated from below, C. chloropygaagain landed on the most brightly illuminated side. In contrast, reducing the brightness on the upper surface of the horizontally-suspended object reduced the number of L. sericataalighting on the upper surface but also reduced the overall catch, suggesting that for L. sericata the response to the illumination of a surface does not outweigh its response to its orientation. The implications of these results for the development of trapping technology for the monitoring or control of these economically important pest species is discussed.