The potential for control of the blowfly Lucilia sericata using odour-baited targets

Abstract A deterministic simulation model is used to explore the potential for control of the sheep blowfly, Lucilia sericata , using odour-baited targets. The simulations show that the ambient temperatures of 15^oC or 20^oC, 13% and 24% of the females present in a population would need to be killed per day, respectively, to prevent the population growing from its initial spring density during the summer blowfly season. In contrast, if both sexes could be sterilized, so that sterilized males were also able to disrupt the fertility of unsterilized females, only 6.5–13% of both sexes would need to be attracted and sterilized at 15^oC or 20^oC respectively. To examine whether these numbers of individuals could be attracted by targets baited with liver and sodium sulphide, mark-release-recapture studies in sheep pastures were used to quantify sticky target efficiencies. The percentage of the Lsericata population caught on any one day was shown to be positively related to average ambient temperature. At 15^oC each sticky target caught approximately 13% and at 20^oC, 41% of the females within a 20 m radius. The results indicate that, at ambient temperatures between 15 and 20^oC, if distributed at a density of approximately 5/ha, targets baited with liver and sodium sulphide could effectively suppress a population of L.sericata. Lower densities of targets would be required if they were able to sterilize both sexes rather than kill or if more attractive synthetic semiochemical baits could be developed.     

Effects of targets impregnated with the chitin synthesis inhibitor triflumuron on the blowfly Lucilia sericata

The effects of exposure of adults of the blowfly Lucilia sericata (Meigen) for 24 h to cloth targets impregnated with the chitin synthesis inhibitor triflumuron plus sucrose or sucrose only, were considered. When both male and female L. sericata were exposed to targets treated with sucrose only, the percentage egg hatch varied between 60–75%. When both sexes were exposed to targets treated with 10% triflumuron plus sucrose the percentage egg hatch varied between 3–7%. When females alone were exposed to triflumuron, prior to the introduction of unexposed males, egg hatch varied between 4–14%. The exposure of males alone to 10% triflumuron treated targets, prior to the introduction of unexposed virgin females, did not give any consistent significant decrease in the mean percentage egg hatch. However, repetition of this trial with males exposed to targets treated with 15 or 20% triflumuron, resulted in a percentage egg hatch of 3.1% and 10.3%, respectively. These results suggest that at relatively high target concentrations, males can transfer effective doses of triflumuron to unexposed virgin females.The exposure of adults to triflumuron treated targets did not significantly increase the mortality of either male or female L. sericata. Furthermore, bioassays showed that there was no significant decline in the ovicidal effectiveness of targets treated with 10% triflumuron and sucrose solution following exposure to sun and rain for up to 8 weeks in 1995 and 1996.The results suggest that targets treated with triflumuron suspension concentrate and sucrose may form the basis of an effective system for the control of L. sericata in the field. However, the relatively high concentrations of triflumuron required and the low efficiency with which males are able to transfer effective doses to unexposed females may diminish one of the main theoretical advantages of using a chitin synthesis inhibitor, such as triflumuron, over a conventional insecticide.     

Development of an attractive target for the sheep blowfly Lucilia sericata

Coloured, odour-baited, sticky targets, designed to catch the sheep blowfly Lucilia sericata Meigen are described and the results of field trials are presented. The number of L. sericata caught by the targets was positively related to target surface area and was dependent on target colour. The performance of the coloured targets is described by a model in which catch is related negatively to reflectivity in the 300-450 nm (ultra-violet/blue) band of the spectrum and positively to the reflectivity in the 450-580 nm (blue-green-yellow) band. Of the Lucilia blowflies caught by the targets, 13-16% were L.sericata and 72-75% of the L.sericata were female. The number of L.sericata caught appeared to be positively related to the availability of sheep over a relatively localized area and showed a marked decline following insecticidal dipping of sheep. The further development of targets and the implications of the results for L.sericata control are discussed.     

Pre-adult mortality in the blowfly Lucilia sericata

Populations of the blowfly, Lucilia sericata (Meigen) (Diptera: Calliphoridae), have a considerable potential for rapid increase; the lifetime reproductive output of each adult female has been estimated to be between 130 and 172 eggs. Nevertheless, in the field, absolute population densities of this species are relatively low. To account for this difference, the levels of mortality affecting the eggs, feeding and wandering larvae and pupae of L. sericata were assessed in the field and laboratory. Percentage egg hatch was dependent on relative humidity with no egg eclosion at humidities below 50%; there was no significant effect of temperature on egg hatch. On infested sheep, the mean mortality of feeding larvae was 53%, but this ranged widely from 0% to 96%. There was no effect of atmospheric temperature or humidity on the mortality of feeding larvae in vivo. In the laboratory, only 10% of wandering larvae pupariated at 10 degrees C. At above 20 degrees C pupariation was consistently almost 100%. Percentage emergence increased from 0% at 10 degrees C to about 80% between 20 and 30 degrees C. The upper lethal temperature for pupae was approached at 35 degrees C. Analysis of the predation of pupae in the field revealed a weak, but significant curvilinear relationship between temperature and proportionate mortality and a median mortality of 0.49% per 24 h exposure (interquartile range = 5.2%). There was no evidence of density dependence in pupal predation. Overall, it is estimated that pre-adult mortality accounts for losses of approximately 97% of each generation, but this figure is subject to considerable variation depending on factors such as climate, time of year and host susceptibility.     

Population suppression for control of the blowfly Lucilia sericata and sheep blowfly strike

Abstract.

• 1 The control of ovine myiasis by suppression of populations of the blowfly Lucifia sericata was investigated experimentally on three farms in the south-west of England in 1992 and 1993.
• 2 In blind trials, sheep on one farm (control) were given two doses of placebo, on a second two doses of the larvicide cyromazine (Vetrazin®, CibaGeigy), and on a third cyromazine and a subsequent dose of placebo.
• 3 The first treatment was given shortly before the predicted spring emergence of L.sericata and the second shortly before the predicted emergence of the second generation. Previous simulation analysis had identified strategic early-season treatment as the optimum for blowfly population suppression.
• 4 On both treatment farms significantly smaller L.sericata populations were recorded throughout 1992 and the incidence of strike was significantly lower than on the control farm. The results show that appropriate early-season timing of sheep treatment can suppress populations of L.sericata and could be used by farmers to reduce the incidence of blowfly strike.
• 5 The results suggest, however, that the effectiveness of population suppression and strike incidence may have been influenced by immigration into the control areas and by adverse weather, the latter changing the susceptibility of sheep to strike and resulting in rising strike incidence even when L.sericata population densities were low. In practice, therefore, blowfly population suppression should be employed as a component of an integrated strike management programme.

     

Antibacterial properties of larval secretions of the blowfly, Lucilia sericata

The antibacterial properties of secretions aseptically collected from larvae of the greenbottle fly Lucilia sericata (Meigen) (Diptera: Calliphoridae) were examined. These investigations revealed the presence of small (<1 kDa) antibacterial factor(s) within the larval secretions, active against a range of bacteria. These include the Gram-positive Staphylococcus aureus, both methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-sensitive Staphylococcus aureus (MSSA), Streptococcus pyogenes and to a lesser extent the Gram-negative Pseudomonas aeruginosa. These secretions were shown to be highly stable as a freeze-dried preparation and, considering the activity against organisms typically associated with clinical infection, may be a source of novel antibiotic-like compounds that may be used for infection control and in the fight against MRSA.     

Effects of ovarian development and protein deprivation on the activity and locomotor responses of the blowfly, Lucilia sericata, to liver odour

Abstract. The spontaneous activity and locomotor responses to liver odour of females of the blowfly Lucilia sericata Meigen (Diptera: Calliphoridae) were examined in the laboratory using a rocking-box olfactometer. Spontaneous activity levels depended on fly age, previous exposure to protein and recent oviposition history; 1-day-old females, 3-day-old protein-fed females, and females that had recently oviposited exhibited lower levels of spontaneous activity than protein-deprived or fully gravid individuals. Changes in locomotor activity in response to liver odour also depended on previous exposure to protein. In protein-fed flies, activity decreased in response to liver odour and then increased on termination of an odour pulse. This response became more pronounced with age, broadly in line with the expected state of oocyte maturation. This pattern of change was not evident in protein-deprived females. The results are discussed in relation to the relative importance of protein and oviposition-site resources to L. sericata and the resultant age and ovarian development-stage biases evident in field catches.     

Suppression of the blowfly Lucilia sericata using odour-baited triflumuron-impregnated targets

Abstract. Field trials were carried out in 1995 and 1996 on farms in the south-west of England to assess the extent to which odour-baited targets could be used to suppress populations of the ectoparasitic blowfly, Lucilia sericata , in sheep pastures. Targets were constructed from 41 × 41 cm squares of aluminium sheet, covered by white cloth which had been dipped in a mixture of sucrose solution (50% w/v) and the chitin synthesis inhibitor triflumuron (10% suspension concentrate). Each target was baited with ≈ 300 g of liver and sodium sulphide solution (10%). Three matched sheep farms were used in the trials. In 1995, triflumuron-impregnated targets were placed around the periphery of sheep pastures at one of the farms in late June, at approximately one target per hectare. In 1996, triflumuron-impregnated targets were placed around the periphery of sheep pastures of a second of the farms in early May, at approximately five targets per hectare. Each year, five sticky targets, used to monitor the L. sericata populations, were also placed in fields at the experimental and the other two farms, which acted as controls. In 1995, the results provided some, although inconclusive, evidence that the triflumuron-impregnated targets had reduced the numbers of L. sericata relative to the populations on the two control farms. In 1996, however, the density of L. sericata on the experimental farm was reduced to almost zero and remained significantly lower than on two control farms throughout the period during which the triflumuron-impregnated targets remained in the field. The results are discussed in relation to the use of triflumuron-treated targets as a practical means of controlling L. sericata and sheep blowfly strike.     

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.     

Visual and olfactory cue interaction in resource-location by the blowfly, Lucilia sericata

Abstract. The interactions between olfactory and visual cues in the landing responses of the blowfly Lucilia sericata (Meigen) (Diptera: Calliphoridae) were examined in a laboratory wind tunnel. The odour of liver and sodium sulphide solution released at 1 L/min, resulted in a greater number of landings, closer to, but downwind of, the central odour release point, than when odour was absent. Three-day-old protein-deprived females landed in greater numbers than protein-fed female flies of the same age; no difference existed between 6-day-old protein-fed or protein-deprived females. Six-day-old, protein-fed flies landed closer to the odour source than did 3-day-old, protein-fed flies. In the presence of odour, flies landed closer to the central release point when it was accompanied by a visual cue. No such effect of the visual cue was evident in the absence of odour. When a plain white sticky-surface (25 × 25 cm) was paired with an identical white surface plus odour, approximately equal numbers of flies landed when the surfaces were placed together; increasingly higher numbers of flies landed on the surface with the odour cue when the distance between the surfaces was increased. When a white surface with a visual cue was presented with the plain white surface plus odour, more flies landed on the white surface with the visual cue than on the plain white surface with odour when they were close together. However, as these two surfaces were moved apart, flies landed increasingly more on the surface with the odour than the surface with the visual cue. The results show that while odour cues may enhance the induction of landing by female L. sericata , visual cues are important when selecting a final landing site.     

The use of carrion as breeding sites by the blowfly Lucilia sericata and other Calliphoridae

Abstract. To examine the species composition of flies breeding in carrion in the field, the carcasses of mice and quail were exposed on sheep farms in the South West of England. Calliphora vicina was the dominant species of Diptera; 19,294 individuals emerged from 175 of the 241 infested carcasses recovered. Lucilia sericata emerged from only 39 of the infested carcasses, at a median of 10 adults per infested carcass. Other species of Lucilia present were L.ampullacea, L.caesar and L.illustris. The highest number of L.sericata emerged from carcasses placed in open pasture, the highest number of C.vicina emerged from carcasses in hedgerow, whereas the highest numbers of L.caesar, L.ampullacea and L.illustris emerged from carcasses in woodland. The duration of exposure of carcasses in the field was negatively related to the size of the adult L.sericata which emerged and, in woodland and hedgerow habitats, to the number of L.sericata which emerged. These data indicate that the larvae of L.sericata in carcasses experience significant levels of competition and that the intensity of this competition may be sufficient to reduce the numbers of L.sericata able to emerge successfully. The size distributions of female L. sericata which emerged from carcasses or which were caught as adults in the field showed only a small degree of overlap, suggesting that only a relatively small proportion of the wild L.sericata population emerge from carcass breeding sites. The results are discussed in relation to the development of new approaches to the control of blowfly strike of sheep.     

Responses of the sheep blowflies Lucilia sericata and Lcuprina to odour and the development of semiochemical baits

Abstract. The literature relating to the attraction of the sheep blowflies Lucilia sericata and Lucilia cuprina to their ovine hosts is reviewed. The responses of the two species are similar and different components of the behaviour leading to host location and oviposition appear to involve at least two distinct sets of semiochemical cues. Activation, upwind orientation and landing appear to occur in response to putrefactive sulphur-rich volatiles, originating from bacterial decomposition products. Oviposition is elicited primarily by the presence of ammonia-rich compounds; moisture, pheromones and tactile stimuli may also act as oviposition stimuli. There is a pronounced sex difference in the response of Lucilia to semiochemicals with a higher proportion of females attracted than males and a higher proportion of gravid than non-gravid females. While the mechanisms of host location by Lucilia are of intrinsic interest, understanding the responses to semiochemicals is important in the attempt to develop powerful synthetic baits for deployment with the traps or targets used for population sampling or suppression. The literature is discussed with respect to the development of synthetic semiochemical baits.     

Mitochondrial genomes of the sheep blowfly, Lucilia sericata, and the secondary blowfly, Chrysomya megacephala

This paper presents complete mitochondrial genomes for the sheep blowfly, Lucilia sericata (Meigen), and the secondary blowfly, Chrysomya megacephala (Fabricius). Both L. sericata and C. megacephala had standard dipteran-type mitochondrial genome architectures and lengths of 15 945 bp and 15 831 bp, respectively. Additionally, C. megacephala possessed a tRNA duplication either side of the D-loop, as previously reported in another Chrysomya species, C. putoria; this duplication appears to be synapomorphic for the genus Chrysomya. As in other insect mitochondrial genomes, base compositions had a high AT content, with both genomes more than 76% AT-rich.     

The effect of bacterially‐dense environments on the development and immune defences of the blowfly Lucilia sericata

Competitive interactions between insects and microbes and the associated cost of development in bacterially‐dense environments are investigated using the blowfly Lucilia sericata (Meigen) (Diptera: Calliphoridae) as a model. The effects of developing in a bacterially‐dense environment are measured by assessing the fitness consequences of competition using the pathogen Staphylococcus aureus. Fitness is quantified in terms of larval survival, puparial development and adult emergence.The influence of bacteria on larval immune defences is investigated using optical density to assess whether antibacterial potency of the larval excretion/secretion changes in response to the degree of contamination of the larval environment. The results obtained demonstrate that bacterial presence has no detrimental effect on survival of L. sericata from egg to adult eclosion, or on puparial size. Additionally, the level of microbial contamination of larvae has no effect on the antibacterial potency of the larval excretion/secretion. These findings confirm that larval antibacterial activity is not induced by the presence of environmental bacteria but is produced constitutively.     

Adult mortality and oviposition rates in field and captive populations of the blowfly Lucilia sericata

Abstract.  1. Adult mortality and oviposition rates were determined for populations of the blowfly Lucilia sericata (Meigen) (Diptera: Calliphoridae). This species is of economic importance as the primary agent of sheep myiasis throughout north-western Europe.
2. Populations of marked flies in six, 1 m³, outdoor field cages and unmarked wild flies at two farms in south-west England were studied simultaneously between May and September 1998.
3. In the field, wild female L. sericata were caught and aged using a combination of wing-fray and ovarian dissection techniques. Survivorship analysis gave estimates of mortality of 1.94% (± 0.037) and 2.09% (± 0.044) per day-degree and mean life expectancy of 51.5 and 47.9 day-degrees above a threshold of 11 °C, at the two farms studied. Mean lifetime reproductive output in the field was estimated to be 159.6 and 138.4 eggs per female at the two farms respectively.
4. The survivorship of cohorts of marked female flies in cages was followed by counting the number of dead individuals each day; the mortality rate of these flies was 0.81% per day-degree (± 3.49 × 10⁻⁴%) and the mean life expectancy was 123.1 day-degrees above a threshold of 11 °C. Mortality rate was shown to increase significantly with average ambient temperature and relative humidity lagged for two sample periods (approximately 10 days). Oviposition rate also increased with average temperature but declined with average relative humidity. A best-fit multiple regression model incorporating both ambient temperature and humidity explained 60.5% of the variance in the pattern of oviposition.
5. The differences between the field and cage populations highlight the caution required when extrapolating life-history parameters from artificial to natural habitats.     

A simple larval diet for population studies on the blowfly Lucilia sericata (Diptera: Calliphoridae)

1. A simple artificial diet was devised for larvae of Lucilia sericata Meigen. 2. A basic diet of 20 g/l agar with 20% horse blood and added yeast sustained normal growth and development, as compared with a lamb meat control. 3. When yeast was added to the blood agar diet at 50 g/l, both peak and final larval weights were increased by 25-50% at higher larval densities (2-8 larvae/g diet) and the hatchling-adult development period was reduced by about 3 days. 4. No adult insects emerged on an agar-yeast diet without blood. Increasing the concentration of blood from 10% to 20% increased adult weight at emergence by 50%, though developmental period was not significantly affected. 5. Increasing larval density significantly reduced the weights of pupae and emergent adults (by up to 50%), both on the blood-agar-yeast diet and the lamb meat control. Percentage survival fell from 80-90% to below 10% on the blood-agar-yeast diet at the highest densities of 4 or 8 larvae/g diet, though survival on lamb was not significantly affected by density.     

Mortality rate, reproductive output, and trap response bias in populations of the blowfly Lucilia sericata

1. The ages of 877 females of the blowfly Lucilia sericata, collected from two farms in south-west England between May and September 1996, were determined using a combination of ovarian dissection and wing fray analysis. 2. Using survivorship analysis, the mortality rates over the entire field season were estimated to be 2.2 and 1.9% per day-degree at the two farms, respectively. These gave a mean life expectancy of 46 and 53 day-degrees, and a lifetime reproductive output of 130 and 172 eggs per female. 3. The mortality rate remained relatively constant over time, with one exception, the first sample of the season when no older flies were present, indicating that underlying changes in demography (emergence and senescence of generations) over the field season were not sufficient to affect the rates calculated by survivorship analysis. 4. The response of L. sericata to liver-baited traps was affected strongly by age and reproductive status, with gravid females and females in the early stages of ovarian development being most markedly over-represented relative to other age categories.     

Antibacterial substances of low molecular weight isolated from the blowfly, Lucilia sericata

Low molecular weight compounds were isolated by high-performance liquid chromatography from the maggot or haemolymph extracts of Lucilia sericata (Meigen) (Diptera: Calliphoridae). Using gas chromatography-mass spectrometry analysis, three compounds were obtained: p-hydroxybenzoic acid (molecular weight 138 Da), p-hydroxyphenylacetic acid (molecular weight 152 Da) and octahydro-dipyrrolo[1,2-a;1',2'-d] pyrazine-5,10-dione (molecular weight 194 Da), also known as the cyclic dimer of proline (or proline diketopiperazine or cyclo[Pro,Pro]). All three molecules revealed antibacterial activity when tested against Micrococcus luteus and/or Pseudomonas aeruginosa, and the effect was even more pronounced when these molecules were tested in combination and caused lysis of these bacteria.     

Induction of antibacterial activity in larvae of the blowfly Lucilia sericata by an infected environment

Maggot debridement therapy (MDT) is a method for the treatment of intractable, infected and necrotic wounds. In MDT, sterile larvae of Lucilia sericata Meigen (Diptera: Calliphoridae) are applied to infected wounds, where they exert antibacterial effects. Once the larvae are placed in the wound, they are no longer germ-free. This study analysed the influence of infected environments on larval antibacterial activities. Sterile larvae were mixed in a test tube containing a bacterial suspension of Staphylococcus aureus or Pseudomonas aeruginosa, transferred to liver puree agar, and incubated at 25 °C for set periods. To collect the larval extracts, the incubated larvae were transferred to a test tube containing phosphate buffered saline (PBS), cut into multiple pieces with scissors, and centrifuged. The supernatant was used to test antibacterial activities. The results showed that infected larvae had better antibacterial capacities than sterile larvae. Antibacterial activities were induced by pretreatment with a single bacterial species, S. aureus or P. aeruginosa, within 24 h and 12 h, respectively, and disappeared after 36 h. The activities were effective against S. aureus, but not against P. aeruginosa. This natural infection model is very similar to the clinical wound context in MDT and will be a powerful tool with which to study the antibacterial activities of L. sericata larvae in MDT.     

Bacteria associated with Lucilia sericata larvae reared on fish wastes

The green blowfly, Lucilia sericata (Meigen) (Diptera: Calliphoridae), is a cosmopolitan species of great medical, veterinary, and forensic importance. In addition, their larvae are among the most promising agents for the bioconversion of low-quality biomass, such as organic waste, into sustainable and nutritionally valuable proteins for farmed fish and poultry. Despite the considerable medical and economic importance, the current literature provides limited information about microbiota associated with larvae. The present study aims to fill this knowledge gap. Freshly harvested L. sericata larvae (maggots) grown on fish wastes were investigated by conventional and molecular approaches to evaluate culturable microbial numbers and unculturable microbial diversity associated with the larval cuticle (external samples) and the internal body. In total 200 bacterial isolates were obtained; 46% of the strains originated from external samples and 58% originated from internal body samples produced extracellular protease enzymes, which may be involved in the digestion of proteins during larval feeding. In total 12 predominant bacteria with high proteolytic activity were further identified by morphological, physiological, biochemical, and molecular tools. Proteolytic bacteria in internal samples included Proteus, Providencia, Micrococcus, Deinococcus, whereas in external samples Providencia, Pseudomonas, and Acinetobacter were found. 16S rRNA clone library analysis revealed that the majority of internal bacteria (35%) were taxonomically assigned as Xanthomonadaceae (Schineria, Xylella, Ignnatzchineria), 28% Morganellaceae (Proteus, Providencia, Serratia), and 14% Enterobacteriaceae (Vagococcus, Serratia). Less abundant were bacteria of the genera Clostridium (3%), Erypelothrix (3%), and Oceanispherum (2%). This knowledge will be useful for biotechnological application of L. sericata.