Host location and behavioural response patterns of the parasitoid,Tachinaephagus zealandicus Ashmead (Hymenoptera: Encyrtidae), to host and host‐habitat odours

Abstract 1. To overcome the challenge of host location in patchy and complex environments, many parasitoids exploit host‐habitat derived odour cues. This study investigated the role of odour cues used during host location by the generalist parasitoid, Tachinaephagus zealandicus, a common parasitoid of Dipteran larvae found in association with decomposing carrion. 2. A Y‐tube olfactometer was used to test aspects of host searching behaviour of T. zealandicus females under different choice scenarios: combinations of host–liver complex, eaten liver, unwashed host larvae, washed host larvae, liver in six stages of decay, and control (no odour). 3. T. zealandicus females demonstrated an innate response to, and preference for, odours arising from liver substrates following interaction with the host. However, this was dependent on the stage of meat decay. Females were not attracted to liver that had never been in contact with a host, regardless of the stage of decay. There were discernible differences in parasitoid movement and behaviour under different choice scenarios. The absence of odour stimulus elicited limited movement about the olfactometer. In the presence of any odour stimuli, females were more active within the olfactometer, and when presented with two odour options T. zealandicus females demonstrate a heightened investigation response, moving across all regions of the olfactometer. 4. Knowledge of the cues that mediate host location for T. zealandicus can contribute to fields such as forensic entomology, where the presence of parasitoids on decomposing remains can be used to estimate time since death.     

Control of the sheep blowfly in Australia and New Zealand – are we there yet?

The last 50 years of research into infections in Australia and New Zealand caused by larvae of the sheep blowfly, Lucilia cuprina, have significantly advanced our understanding of this blowfly and its primary host, the sheep. However, apart from some highly effective drugs it could be argued that no new control methodologies have resulted. This review addresses the major areas of sheep blowfly research over this period describing the significant outcomes and analyses, and what is still required to produce new commercial control technologies. The use of drugs against this fly species has been very successful but resistance has developed to almost all current compounds. Integrated pest management is becoming basic to control, especially in the absence of mulesing, and has clearly benefited from computer-aided technologies. Biological control has more challenges but natural and perhaps transformed biopesticides offer possibilities for the future. Experimental vaccines have been developed but require further analysis of antigens and formulations to boost protection. Genetic technologies may provide potential for long-term control through more rapid indirect selection of sheep less prone to flystrike. Finally in the future, genetic analysis of the fly may allow suppression and perhaps eradication of blowfly populations or identification of new and more viable targets for drug and vaccine intervention. Clearly all these areas of research offer potential new controls but commercial development is perhaps inhibited by the success of current chemical insecticides and certainly requires a significant additional injection of resources.     

Microsporidiosis of Tachinaephagus zealandicus Ashmead (Hymenoptera: Encyrtidae)

An undescribed microsporidium was found infecting Tachinaephagus zealandicus, a gregarious parasitoid that attacks third instar larvae of muscoid flies. Spores were present in all body regions and in all stages of development. Infected adults contained an average of 3.75 x 10(5) spores, and the pathogen was vertically transmitted to progeny. Infected female adults were fed either rifampicin or albendazole mixed with honey to determine the effectiveness of these drugs in preventing vertical transmission. After eight days of feeding on rifampicin the parasitoids produced progeny of which only 37% were infected. In contrast, albendazole-treated and untreated females produced progeny that were 97% and 100% infected, respectively. Healthy and infected colonies were established and studies were conducted to determine the mechanisms of transmission. It was observed that the efficiency of vertical (maternal) transmission was 96.3%. Uninfected parasitoid immatures also became infected when they shared superparasitized hosts with infected immatures. The method of transmission within superparasitized hosts is not known.     

Progress towards the development of a transgenic strain of the Australian sheep blowfly (Lucilia cuprina) suitable for a male-only sterile release program

The Australian sheep blowfly Lucilia cuprina is the most important pest species involved in cutaneous myiasis (flystrike) of sheep in Australia and New Zealand. In New Zealand L. cuprina is primarily controlled through the application of insecticides. However, there is an increased interest in biological methods of control of this species. We have proposed to develop a genetically modified strain of L. cuprina that would be ideal for a male-only sterile release program. To that end we have developed a method for making transgenic L. cuprina using a piggyBac vector and an EGFP marker gene. We have also developed in Drosophila melanogaster a 2-component genetic system for controlling female viability. Females carrying both components of the system die unless fed a diet that contains tetracycline. We anticipate that the female-killing system will need to be optimised for L. cuprina in order to make a strain with the properties required for a male-only release program.     

Habitat preferences and carcase colonization by sheep blowflies in the northern North Island of New Zealand

Abstract. In urban Auckland, from September 1991 to May 1992, only six specimens of the Australian sheep blowfly, Lucilia cuprina, were collected in farmed parkland and in garden habitats. The common green blowfly, Lucilia sericata, was the most common calliphorid trapped in these habitats. Neither of the two Lucilia species were found in native bush remnants in urban or rural areas where Calliphora hilli was dominant. Similarly very few L.cuprina (0.3% of the total) were trapped in rural rubbish tips in January and February where the majority of blowflies were again L.sericata. These results are compared with data collected from pastoral habitats, where L.cuprina is the major cause of flystrike. Lucilia cuprina was reared from five out of ninety-nine carcases found in rural areas. Calliphora stygia and L.sericata were the most common blowflies colonizing carrion and were reared from 59% and 51% of the carcases respectively.     

Estimating the incidence of fly myiases in Australian sheep flocks: development of a weather-driven regression model

The blowfly, Lucilia cuprina Wiedemann (Diptera: Calliphoridae), is the primary myiasis (strike) fly of sheep in Australia. Most strike occurs in the anal-perineum area (crutch), but strike to the neck, shoulders, back and withers (body) is also important. Regression analysis was used to determine the extent to which the weekly incidence of flystrike can be explained by variations in fly abundance and/or recent changes in weather, pasture conditions or flock management. Strike and flock management data were collected by questionnaire surveys of 30-60 sheep properties in each of three major sheep-producing areas in southeastern Australia, namely, Gunning (southern New South Wales), Inverell (northern New South Wales) and Flinders Island (Bass Strait). After using simulation modelling to remove effects due to shearing, crutching and/or insecticide treatment, pasture growth index was found to be the most important explanatory variable affecting the incidence of all forms of myiasis. Others were average weekly air temperature, the amount and frequency of rainfall, relative humidity, dung quality index and a factor denoting seasonal effects. Together, these variables accounted for 48.4% of the variation in body strike, 56.8% of that in crutch strike and 51.9% of that in other forms of strike. Prediction was improved by the inclusion of additional lagged variables describing previous strike, fly abundance and fly activity. With these additions, the variation explained increased to 60.4% for body strike, 68.0% for crutch strike and 58.3% for other strikes.     

Efficacy of native and recombinant Cry1B protein against experimentally induced and naturally acquired ovine myiasis (fly strike) in sheep

Several hundred strains of Bacillus thuringiensis (Bt), isolated in New Zealand from samples of soil and sheep fleece, were tested for toxicity to larvae of the blowfly Lucilia cuprina Wiedemann. Characterization of the Bt strains revealed that three of the more active strains produced Cry1Ba (an insecticidal protein present in Bt mother cell crystal inclusion) that was toxic to blowflies. These strains were evaluated for the ability to prevent experimentally induced fly strike in a bioassay by using first instars. Results with undiluted spore/crystal preparations were variable, but they generally prevented fly strike on sheep maintained on pasture for 3-6 wk. Spore viability was satisfactory throughout the trials and environmental factors (e.g., precipitation and UV radiation) seemed to have minimal effect on persistence. The loss of fly strike protection in these experiments correlated with the movement of spore/crystal toxicity away from the skin as a result of wool growth. Solubilized protein preparations were not as potent as spore/crystal preparations and fly strike protection lasted only from 1 to 3 wk. Vegetative forms of the Cry1Ba-producing strains of Bt did not establish on the fleece of sheep, did not produce significant sporulation, and no protection against fly strike was achieved. Escherichia coli expressing recombinant Cry1Ba protein was toxic to larvae in vitro but did not effectively protect sheep from fly strike because blowfly larvae were able to establish readily 8 d posttreatment. In a single field experiment involving 80 sheep per group, a spore/crystal preparation from a Bt strain expressing Cry1Ba provided less protection from naturally acquired fly strike than afforded by a commercially available dip.     

Delaying insect access alters community composition on small carrion: a quantitative approach

We present a study of interactions in the highly competitive insect communities inhabiting the carrion of small mammals. Via manipulation in a fully quantitative design, we delayed community development by excluding insect colonization in mouse and rat carcasses for 3 days, to study the role of early competitively dominant colonizers [burying beetles (Nicrophorus spp.; Coleoptera: Silphidae) and blowfly larvae (Diptera: Calliphoridae)] in the course of heterotrophic succession on small cadavers. Earlier studies demonstrated that in the case of large mammalian carrion, exclusion of insects’ access to the carcass in the early stages of decomposition altered the successional trajectory and species assemblages. However, the effect of such manipulation in easy monopolizable small vertebrate carrion remained unknown. Our results demonstrate that delaying insect access to carrion significantly lowered blowfly larvae abundances, while it simultaneously had no effect on colonization and carrion burial by burying beetles. Higher abundances of blowfly larvae seem to deter necrophagous beetles, whereas they are not harmful to the larvae of flesh flies, at least in larger rat carcasses. Predatory beetle species preferred the lower abundances of blowfly larvae, presumably due to better accessibility of their prey. Our results therefore suggest that in the course of the entire season, larvae of blowflies are the dominant competitors in small carcasses, and significantly affect the assembly of other insect groups, whereas burying beetles may exhibit a more temporal pattern of dominance.     

A new host record forTachinaephagus zealandicus [Hym.: Encyrtidae]

Tachinaephagus zealandicus Ashmead was reared from puparia ofProtocalliphora n. sp. [Dip.], representing a new host record for this parasitoid. The habitat ofProtocalliphora is qualitatively different than those of all previously recorded hosts for this parasitoid.T. zealandicus has been imported from Australia and New Zealand and this record strengthens the belief that it has become established in California.     

Virus-like particles in the ovaries of microctonus aethiopoides loan (Hymenoptera: braconidae), a parasitoid of adult weevils (Coleoptera: curculionidae)

The morphology of the female reproductive system of Microctonus aethiopoides is described and illustrated, and an ultrastructural examination of the ovaries was carried out. Virus-like particles (VLPs) were initially found in the ovarial epithelial cells of females from pre-adult emergence from the pupal cocoon until at least 5 days after emergence. The particles assembled in the nucleus of the epithelial cells, apparently being synthesized de novo in association with a putative virogenic stroma, and they moved into the lumen of the ovarioles surrounding the developing eggs. VLPs were also found in some other cells in both male and female M. aethiopoides. VLPs have not been found in M. hyperodae or the New Zealand native species M. zealandicus. The function of the VLP and its possible role in potential parasitoid host range determination are discussed. Copyright 1999 Academic Press.     

Insect growth regulator cross-resistance studies in field- and laboratory-selected strains of the Australian sheep blowfly, Lucilia cuprina (Wiedemann) (Diptera: Calliphoridae)

Abstract Resistance to diflubenzuron in the Australian sheep blowfly, Lucilia cuprina , has rendered this insecticide incapable of preventing flystrike in sheep from a few districts in eastern Australia. Wool producers affected by this situation must find suitable alternatives to protect their flocks. Results of laboratory bioassays against one population demonstrated that, despite extremely high diflubenzuron resistance (Resistance Factor >791), it had only very low (2x) tolerance of cyromazine and dicyclanil. It is unlikely that this level of tolerance would have any practical impact on field control with either insecticide. Consequently, wool producers in districts where diflubenzuron-resistant flies are common can rotate insecticide treatment to either of these compounds to prevent flystrike in their flocks. However, unlike the highly diflubenzuron-resistant field strain, a laboratory strain selected for resistance to diflubenzuron (Resistance Factor = 617) was 10 times more resistant to dicyclanil than a susceptible strain but, like the field strain, was only two times more tolerant of cyromazine. Conversely, a field-derived strain selected in the laboratory for cyromazine resistance was 20 times more resistant to dicyclanil and 362 times more resistant to diflubenzuron than the reference susceptible strain.     

Direct and indirect top‐down effects of previous contact with an enemy on the feeding behavior of blowfly larvae

We investigated the addition of a trophic level to a simple food web. Direct and indirect effects caused by the presence of a new species in the food web were quantified by estimating survival and consumption rates on the basal resource. We focused on a blowfly intraguild prey–predator system with various ecological interactions taking place during the larval period. The experiments were designed to set Chrysomya megacephala (Fabricius) (Diptera: Calliphoridae) as the intraguild prey and Chrysomya albiceps (Wiedemann) as the intraguild predator and/or cannibal. The generalist pupal parasitoid Nasonia vitripennis (Walker) (Hymenoptera: Pteromalidae) was introduced into the system during a non‐susceptible life stage of the interacting blowfly species. The cascading parasitoid effects induced behavioral changes in the blowfly larvae, increasing the impact of intraguild predation and cannibalism on blowfly survival. The results suggest that blowfly larvae can change their feeding behavior in response to the presence of a parasitoid.     

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.     

Mitochondrial phylogeography of New Zealand freshwater crayfishes, Paranephrops spp

Tectonic movement at the boundary of the Indo-Australian and Pacific Plates during the Miocene and Pliocene is recognized as a driving force for invertebrate speciation in New Zealand. Two endemic freshwater crayfish (koura) species, Paranephrops planifrons White 1842 and Paranephrops zealandicus White 1842, represent good model taxa to test geological hypotheses because, due to their low dispersal capacity and life history, geographical restriction of populations may be caused by vicariant processes. Analysis of a mitochondrial DNA marker (cytochrome oxidase subunit I) reveals not two, but three major koura lineages. Contrary to expectation, the cryptic West Coast group appears to be more closely related to P. zealandicus than to P. planifrons and has diverged earlier than the final development (Late Pleistocene) of Cook Strait. Our date estimates suggest that koura lineage diversification probably coincided with early to mid-Alpine orogeny in the mid-Pliocene. Estimates of node ages and the phylogenies are inconsistent with both ancient Oligocene and recent postglacial Pleistocene range expansion, but suggest central to north colonization of North Island and west to east movement in South Island during mid- to late Pliocene. Crypsis and paraphyly of the West Coast group suggest that morphological characters presently used to classify koura species could be misleading.     

Parasitism of the lucerne pest Sitona discoideus Gyllenhal (Coleoptera: Curculionidae) and non-target weevils by Microctonus aethiopoides Loan (Hymenoptera: Braconidae) in south-eastern Australia, with an assessment of the taxonomic affinities of non-target hosts of M. aethiopoides recorded from Australia and New Zealand

Abstract  The braconid parasitoid Microctonus aethiopoides Loan has been released in Australia and New Zealand for biological control of the lucerne pest Sitona discoideus Gyllenhal. In New Zealand, the parasitoid attacks a number of endemic weevil species. A survey of Curculionoidea found in and near lucerne in south-eastern Australia was carried out to investigate whether similar non-target parasitism was occurring, and to relate this to levels of parasitism found in the target host, S. discoideus . Some of the original M. aethiopoides release sites were particularly targeted in the survey of 25 sites in Victoria, New South Wales and South Australia. Almost 2500 weevils were collected, of which over 90% were S. discoideus , with the remaining 197 other weevils comprising 29 species found at 15 of the 25 sites. Parasitism of S. discoideus by M. aethiopoides occurred at 12 lucerne sites, with levels ranging from 0 to 25%. A single incidence of parasitism of a species of an Australian native weevil Prosayleus sp. by M. aethiopoides was recorded. No parasitism of any other weevil species was observed. The taxonomic affinities between Sitona and native Australian and New Zealand weevils are discussed, concluding that non-target host range in M. aethiopoides may be determined more by ecological factors than by taxonomic affinities among its hosts.     

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     

Fly abundance and climate as determinants of sheep blowfly strike incidence in southwest England

The relationships between abundance of the blowfly Lucilia sericata (Meigen) (Diptera: Calliphoridae), climate, animal management procedures and the incidence of cutaneous myiasis (blowfly strike) in sheep were examined in three sheep pasture systems in southwest England during the summers of 2002 and 2003. In each year, flies were collected using liver-baited sticky targets, daily weather and routine husbandry practices were noted and the age-class of each animal infested and body position of each strike were recorded. On sites where no strike control was used, 5.8-12.1% of ewes and 5.7-15.8% of lambs were struck. Ewe strikes predominated at the beginning of the season. The incidence of ewe strikes was significantly associated with higher mean temperature, rainfall and shearing; shearing was associated with a 95% reduction in the risk of ewe strike. In lambs, the incidence of strike was significantly related to higher fly abundance, ewe shearing, treatment and mean ambient temperature. Lambs were 4.6 times more likely to be struck after the ewes had been shorn than before; however, the strongest relationship was with mean L. sericata abundance. Average minimum threshold temperatures of 9.5 degrees C for lamb strikes and 8.5 degrees C for all strikes were extrapolated, below which oviposition did not occur. Over 80% of ewe strikes occurred in the breech region in 2002, as did 100% in 2003. However, in lambs both body and breech strikes occurred in both years. The distribution of lamb strikes appeared to change over time, with breech strikes predominating in May, June and July and body strikes occurring increasingly later in the season. The incidence of lamb breech strikes was significantly associated with higher L. sericata abundance and ewe shearing but there was no relationship with weather conditions. By contrast, the risk of body strike in lambs was significantly associated with higher blowfly abundance, higher rainfall and higher maximum temperatures. The relationship between strike incidence and L. sericata abundance is important because it enhances our understanding of strike incidence patterns and management of this disease. Clearly, any factors that facilitate larger L. sericata populations, such as inappropriate carcass burial or increased average ambient temperatures, are likely to increase the incidence of strike.     

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.     

The seasonal pattern of sheep blowfly strike in England and Wales

Abstract. Information on 4049 cases of sheep blowfly strike on 495 farms in England and Wales was gathered in a longitudinal survey conducted in 1991. The breech was the most commonly infested area, accounting for 70.9% of all strikes recorded. Body strikes accounted for 19.7% and foot strikes 11.4% of all strikes. Early in the season the incidences of both breech and body strikes were higher in ewes than in lambs, but from June to October lamb strikes predominated. The incidence of foot strike was greater in ewes than lambs throughout the year, with a peak incidence in September. Regional differences in strike incidence were evident; the highest overall incidences were recorded in the southern regions of England where peak incidence occurred earlier in the year. The highest monthly incidence risk was recorded in August in the south-east of England where there were 14.2 strikes per 1000 lambs and 4.0 strikes per 1000 ewes at risk. The importance of faecal soiling and ambient temperature are discussed.