Two new species and a new Australian record of Bactrocera Macquart (Diptera : Tephritidae : Dacinae) from Northern Queensland, Torres Strait and Papua New Guinea

Bactrocera (Bactrocera) speewahensis Fay & Hancock sp. n. is described from northern Queensland and B. (B.) torresiae Huxham & Hancock sp. n. is described from Boigu, Saibai and Dauan islands in Torres Strait and southern Papua New Guinea. Bactrocera (B.) nigrovittata Drew is newly recorded from Australia. All records are of male flies responding to chemical lures.

Suppression of populations of Australian sheep blowfly, Lucilia cuprina (Wiedemann) (Diptera: Calliphoridae), with a novel blowfly trap

The Australian sheep blowfly, Lucilia cuprina initiates more than 85% of fly strikes on sheep in Australia with an estimated average annual cost of A$280 million to the Australian wool industry. LuciTrap® is a commercially available, selective trap for L. cuprina consisting of a plastic bucket with multiple fly entry cones and a synthetic attractant. The impact of LuciTrap on populations of L. cuprina on sheep properties in five Australian states was evaluated by comparing L. cuprina populations on paired properties with and without LuciTraps over seasons when significant fly populations could be expected. Twenty-four comparisons (trials) were conducted over four years. During times of ‘higher fly density’ (when the 48 h geometric mean of trap catches on the control property was greater than five L. cuprina), the overall geometric mean trap catches for control and trapped properties differed significantly (P<0.001) with mean trap catches of 19.4 and 7.74 L. cuprina respectively. The selectivity of the LuciTrap was confirmed with 59% of all trapped flies being L. cuprina. Chrysomya spp. and Calliphora spp. constituted 9.3% and 1.1% of the catches with a variety of other flies (mainly Sarcophagidae and Muscidae) providing the remainder (31%). L. sericata was only trapped in Tasmania and made up 7.7% of the Lucilia spp. catch in this State. Seventy-two percent of the trapped L. cuprina were female. The deployment of LuciTrap on sheep properties at one trap per 100 sheep from the beginning of the anticipated fly season suppressed the populations of L. cuprina by 60% compared to matched control properties. The LuciTrap is a selective and easy to use fly trap and constitutes an effective, non-insecticidal tool for use in integrated management programs for L. cuprina.

Specific detection of the Old World screwworm fly, Chrysomya bezziana, in bulk fly trap catches using real-time PCR.

The Old World screwworm fly (OWS), Chrysomya bezziana Villeneuve (Diptera: Calliphoridae), is a myiasis-causing blowfly of major concern for both animals and humans. Surveillance traps are used in several countries for early detection of incursions and to monitor control strategies. Examination of surveillance trap catches is time-consuming and is complicated by the presence of morphologically similar flies that are difficult to differentiate from Ch. bezziana, especially when the condition of specimens is poor. A molecular-based method to confirm or refute the presence of Ch. bezziana in trap catches would greatly simplify monitoring programmes. A species-specific real-time polymerase chain reaction (PCR) assay was designed to target the ribosomal DNA internal transcribed spacer 1 (rDNA ITS1) of Ch. bezziana. The assay uses both species-specific primers and an OWS-specific Taqman MGB probe. Specificity was confirmed against morphologically similar and related Chrysomya and Cochliomyia species. An optimal extraction protocol was developed to process trap catches of up to 1000 flies and the assay is sensitive enough to detect one Ch. bezziana in a sample of 1000 non-target species. Blind testing of 29 trap catches from Australia and Malaysia detected Ch. bezziana with 100% accuracy. The probability of detecting OWS in a trap catch of 50 000 flies when the OWS population prevalence is low (one in 1000 flies) is 63.6% for one extraction. For three extractions (3000 flies), the probability of detection increases to 95.5%. The real-time PCR assay, used in conjunction with morphology, will greatly increase screening capabilities in surveillance areas where OWS prevalence is low.

Function and regulation of Cdk7

Understanding the process of cell division is crucial for modern cancer medicine due to the central role of uncontrolled cell division in this disease. Cancer involves unrestrained proliferation as a result of cells loosing normal control and being driven through the cell cycle, where they normally would be non-dividing or quiescent. Progression through the cell cycle is thought to be dependent on the sequential activation of cyclin-dependent kinases (Cdks). The full activation of Cdks requires the phosphorylation of a conserved residue (threonine-160 on human Cdk2) on the T-loop of the kinase domain. In metazoan species, a trimeric complex consisting of Cdk7, cyclin H and Mat1 has been suggested to be the T-loop kinase of several Cdks. In addition, Cdk7 have also been implicated in the regulation of transcription. Cdk7, cyclin H, and Mat1 can be found as subunits of general transcription factor TFIIH. Cdk7, in this context, phosphorylates the Carboxy-terminal domain (CTD) of the large subunit of RNA polymerase II (RNA pol II), specifically on serine-5 residues of the CTD repeat. The regulation of Cdk7 in these and other functions is not well known and the unambiguous characterization of the in vivo role of Cdk7 in both T-loop activation and CTD serine-5 phosphorylation has proved challenging. In this study, the fission yeast Cdk7-cyclin H homologous complex, Mcs6-Mcs2, is identified as the in vivo T-loop kinase of Cdk1(Cdc2). It also identifies multiple levels of regulation of Mcs6 kinase activity, i.e. association with Pmh1, a novel fission yeast protein that is the apparent homolog of metazoan Mat1, and T-loop phosphorylation of Mcs6, mediated by Csk1, a monomeric T-loop kinase with similarity to Cak1 of budding yeast. In addition, Skp1, a component of the SCF (Skp1-Cullin-F box protein) ubiquitin ligase is identified by its interactions with Mcs2 and Pmh1. The Skp1 association with Mcs2 and Pmh1 is however SCF independent and does not involve proteolytic degradation but may reflect a novel mechanism to modulate the activity or complex assembly of Mcs6. In addition to Cdk7, also Cdk8 has been shown to have CTD serine-5 kinase activity in vitro. Cdk8 is not essential in yeast but has been shown to function as a transcriptional regulator. The function of Cdk8 is unknown in flies and mammals. This prompted the investigation of murine Cdk8 and its potential role as a redundant CTD serine-5 kinase. We find that Cdk8 is required for development prior to implantation, at a time that is co-incident with a burst of Cdk8 expression during normal development. The results does not support a role of Cdk8 as a serine-5 CTD kinase in vivo but rather shows an unexpected requirement for Cdk8, early in mammalian development. The results presented in this thesis extends our current knowledge of the regulation of the cell cycle by characterizing the function of two distinct cell cycle regulating T-loop kinases, including the unambiguous identification of Mcs6, the fission yeast Cdk7 homolog, as the T-loop kinase of Cdk1. The results also indicate that the function of Mcs6 is conserved from fission yeast to human Cdk7 and suggests novel mechanisms by which the distinct functions of Cdk7 and Mcs6 could be regulated. These findings are important for our understanding of how progression of the cell cycle and proper transcription is controlled, during normal development and tissue homeostasis but also under condition where cells have escaped these control mechanisms e.g. cancer.

Controlling fly strike and louse infections in sheep with tea tree oil

This study aimed to assess the effect of tea tree oil based formulations against two major ectoparasitic diseases in the sheep industry, flystrike and louse infestation, and to provide data to assist the assessment of the commercial feasibility of development of tea tree oil based ectoparasiticides. The results demonstrate insecticidal effects against both sheep lice and blowflies and repellent effects against adult flies and maggots. Dipping sheep in a Tea Tree Oil based formulation appeared to completely eradicate lice and suggests its potential use in sheep dipping formulations. Repellent and insecticidal effects against sheep blowflies, together with previously reported anti-microbial and wound healing properties, suggest significant benefits from the inclusion of tea tree oil in flystrike and wound treatment formulations. These effects occurred at concentrations of Tea Tree Oil that suggest the commercial viability of development of Tea Tree Oil based formulations for sheep parasite control and wound treatment and a potential new market for Tea Tree Oil.

Nuisance fly management on cattle feedlots

The Project will assess the impact of chemical applications and more targeted cleaning on nuisance fly control in feedlots. It will develop and evaluate new, biological agents for the control of nuisance flies and explore options for integration of these.

Fungal Biopesticide for cattle tick and buffalo fly control

Cattle ticks and buffalo flies impose significant economic burdens on the Northern Australian cattle and dairy industries. With the increased temperatures expected under climate change the range of parasites such as these is likely to extend. Current control options for these ectoparasites are limited by problems associated with chemical resistance and residues. Fungal biopesticides offer a sustainable and promising alternative method of control. Laboratory and animal studies have established the potential for the fungus Metarhizium in tick control and provided data that suggests a secondary effect of buffalo fly control is possible. Small field trials are required to obtain a proof of concept for the control of ticks and buffalo flies on animals.

CRC20093 Increasing Diagnostic Capacity in Thailand

Accurate identification of pests is essential for practically all aspects of agricultural development and is critical to the operations of biosecurity that safeguard agricultural integrity and facilitate trade. Diagnostic capability is at the forefront of and complementary to, activities such as border protection, incursion management, surveillance and pest and disease certification. The efficiency of a biosecurity system therefore depends largely on the feedback between these activities and diagnostics. Australian scientists will train Thai scientists in diagnostics and surveillance to provide the Thai DOA with skills that will aid in the development of a Thai Diagnostic Network. The skills will be taught using a range of pests, including some which have particular biosecurity importance for both Australia and Thailand such as citrus canker, potato viruses and fruit flies.

Spatial and diurnal pattern of protein foraging by Bactrocera tryoni (Froggatt) on a host plant

Background: Queensland fruit fly, Bactrocera tryoni, is the major pest fruit fly in Australia. Protein bait sprays, where insecticides are mixed with spot applications of a protein based food lure, are one of the sustainable pre-harvest fruit fly management strategies used in Australia. Although protein bait sprays do manage fruit fly infestation in the field, there is little science underpinning this technique and so improving its efficacy is difficult. Lacking information includes where and when to apply protein bait in order to best target foraging B. tryoni. As part of new work in this area, we investigated the effect of height of protein on tree and host plant fruiting status on the spatial and temporal protein foraging patterns of B. tryoni. MEthod: The work was conducted in the field using nectarine and guava plants and wild B. tryoni at Redland Bay, Queensland, Australia. Spot sprays of protein bait were applied to the foliage of randomly selected fruiting and non-fruiting trees. Each tree received protein bait spot sprays on the lower and higher foliage at 0530hrs. The number, sex and species of flies that fed on each protein spot were recorded hourly from 0600hrs through to 1800hrs.Results: For nectarines, there was a significant difference in the number of B. tryoni feeding on protein bait placed at different locations within the tree (ANOVA, F = 8.898, p = 0.001). More flies fed on protein placed on higher foliage relative to lower, irrespective of the fruiting status of the nectarine trees. A significant difference was also observed in the diurnal protein feeding pattern of B. tryoni (ANOVA, F = 2.164, p = 0.024), with more flies feeding at 1600hrs. Results for guava are still being collected and will be presented at the meeting.Conclusions: We conclude that B. tryoni effectively forages for protein at heights higher than 1.3m from ground, indicating greater efficacy of protein bait when applied at foliage higher in the canopy. Bactrocera tryoni actively forages for protein throughout the day, with a highest feeding peak at 1600hrs. The lack of significant difference in the spatial protein foraging pattern between fruiting and non-fruiting nectarine trees may be a real result, or may have resulted from the fruiting tree being very close (within 1 – 2 metres) of the non-fruiting tree. This hypothesis is being tested in the guava trial.

Foraging behaviours of Diachasmimorpha kraussii (Fullaway) (Hymenoptera: Braconidae) and its host Bactrocera tryoni (Froggatt) (Diptera: Tephritidae) in a nectarine (Prunus persica (L.) Batsch var. nectarina (Aiton) Maxim) orchard

Diachasmimorpha kraussii is a larval parasitoid of dacine fruit flies. Host utilisation behaviour, including field foraging behaviour, is poorly known in this species. The diurnal foraging behaviour of D. kraussii and one of its common hosts, Bactrocera tryoni, in a nectarine orchard was concurrently recorded. Observations of mating, resting, feeding and oviposition were taken two-hourly on 42 trees, commencing at 07:00 h and terminating at 17:30 h, for 17 days. Resting and oviposition were common events within the orchard for both species, while mating behaviours were not recorded in the orchard for either species. Feeding was not observed for D. kraussii and was rare for B. tryoni. At the level of the individual tree there was a very weak, but significant correlation between parasitoid and fly abundance over a day, but when broken down to the individual observation periods the correlations were absent, or were weakly significant in an inconsistent manner (i.e. sometimes positively correlated, sometimes negatively correlated). At the orchard level, abundance of the parasitoid was not correlated with adult fly abundance. Results suggest that D. kraussii forage independently to adult B. tryoni, a result consistent with a prediction that their foraging is largely driven by larval or plant damage cues.

Preference among four Bactrocera species (Diptera: Tephritidae) by Diachasmimorpha kraussii (Fullaway) (Hymenoptera: Braconidae)

Diachasmimorpha kraussii is an endoparasitoid of larval dacine fruit flies. To date, the only host preference study done on D. kraussii has used fruit flies from outside its native range (Australia, Papua New Guinea, Solomon Islands). In contrast, this paper investigates host preference for four fly species (Bactrocera cacuminata, Bactrocera cucumis, Bactrocera jarvisi and Bactrocera tryoni), which occur sympatrically with the wasp in the Australian component of the native range. D. kraussii oviposition preference, host suitability (parasitism rate, number of progeny, sex ratio) and offspring performance measures (body length, hind tibial length, developmental time) were investigated with respect to the four fly species in the laboratory in both no-choice and choice situations. The parasitoid accepted all four fruit fly species for oviposition in both no-choice and choice tests; however, adult wasps only emerged from B. jarvisi and B. tryoni. Through dissection, it was demonstrated that parasitoid eggs were encapsulated in both B. cacuminata and B. cucumis. Between the two suitable hosts, measurements of oviposition preference, host suitability and offspring performance measurements either did not vary significantly or varied in an inconsistent manner. Based on our results, and a related study by other authors, we conclude that D. krausii, at the point of oviposition, cannot discriminate between physiologically suitable and unsuitable hosts.

Preference and performance of Diachasmimorpha kraussii (Fullaway) (Hymenoptera: Braconidae) on five commercial fruit species

Diachasmimorpha kraussii is a polyphagous endoparasitoid of dacine fruit flies. The fruit fly hosts of D. krausii, in turn, attack a wide range of fruits and vegetables. The role that fruits play in host selection behaviour of D. kraussii has not been previously investigated. This study examines fruit preference of D. kraussii through a laboratory choice-test trial and field fruit sampling. In the laboratory trial, oviposition preference and offspring performance measures (sex ratio, developmental time, body length, hind tibial length) of D. kraussii were investigated with respect to five fruit species [Psidium guajava L. (guava), Prunis persica L. (peach), Malus domestica Borkh. (apple), Pyrus communis L. (pear) and Citrus sinensis L. (orange)], and two fruit fly species (Bactrocera jarvisi and B. tryoni). Diachasmimorpha kraussii responded to infested fruit of all fruit types in both choice and no-choice tests, but showed stronger preference for guava and peach in the choice tests irrespective of the species of fly larvae within the fruit. The wasp did not respond to uninfested fruit. The offspring performance measures differed in a non-consistent fashion between the fruit types, but generally wasp offspring performed better in guava, peach and orange. The offspring sex ratio, except for one fruit/fly combination (B. jarvisi in apple), was always female biased. The combined results suggest that of the five fruits tested, guava and peach are the best fruit substrates for D. krausii. Field sampling indicated a non-random use of available, fruit fly infested fruit by D. kraussii. Fruit fly maggots within two fruit species, Plachonia careya and Terminalia catappa, had disproportionately higher levels of D. krausii parasitism than would be expected based on the proportion of different infested fruit species sampled, or levels of fruit fly infestation within those fruit.

Evaluation of exclusion netting for insect pest control and fruit quality enhancement in tree crops

This work evaluated the following aspects of the use of exclusion netting in low chill stone fruit: the efficacy of protection from fruit fly for this highly susceptible crop; the effects on environmental factors; and the effects on crop development. Concurrently, an economic viability study on the use of exclusion netting was undertaken. The trial site was a 0.6-ha block of low chill stone fruit at Nambour, south-east Queensland, Australia. In this area, populations of Queensland fruit fly (Bactrocera tryoni) are known to be substantial, particularly in spring and summer. The trial block contained healthy 4-year-old trees as follows: 96 peach trees (Prunus persica cv. Flordaprince) and 80 nectarine trees (40 P. persica var. nucipersica cv. White Satin and 40 P. persica var. nucipersica cv. Sunwright). Exclusion netting was installed over approximately half of the block in february 2001. The net was a UV-stabilized structural knitted fabric made from high-density polyethylene yarn with a 10-year prorated UV degradation warranty. The results demonstrated the efficacy of exclusion netting in the control of fruit flies. Exclusion netting increased maximum temperatures by 4.4 deg C and decreased minimum temperatures by 0.5 deg C. Although exclusion netting reduced irradiance by approximately 20%, it enhanced fruit development by 7-10 days and improved fruit quality by increasing sugar concentration by 20-30% and colour intensity by 20%.

Host Susceptibility of Citrus Cultivars to Queensland Fruit Fly (Diptera: Tephritidae)

Citrus crops are considered to be relatively poor hosts for Queensland fruit fly, Bactrocera tryoni (Froggatt), as for other tephritid species. Australian citrus growers and crop consultants have reported observable differences in susceptibility of different citrus cultivars under commercial growing conditions. In this study we conducted laboratory tests and field surveys to determine susceptibility to B. tryoni of six citrus cultivars [(Eureka lemon (Citrus limon (L.) Osbeck); Navel and Valencia oranges (C. sinensis (L.) Osbeck); and Imperial, Ellendale, and Murcott mandarins (C. reticulata Blanco)]. The host susceptibility of these citrus cultivars was quantified by a Host Susceptibility Index, which is defined as the number of adult flies produced per gram of fruit infested at a calculated rate of one egg per gram of fruit. The HSI was ranked as Murcott (0.083) > Imperial (0.052) ≥ Navel (0.026) ≥ Ellendale (0.020) > Valencia (0.008) ≥ Eureka (yellow) (0.002) > Eureka (green) (0). Results of the laboratory study were in agreement with the level of field infestation in the four citrus cultivars (Eureka lemon, Imperial, Ellendale, and Murcott mandarins) that were surveyed from commercial orchards under baiting treatments against fruit flies in the Central Burnett district of Queensland. Field surveys of citrus hosts from the habitats not subject to fruit fly management showed that the numbers of fruit flies produced per gram of fruit were much lower, compared with the more susceptible noncitrus hosts, such as guava (Psidium guajava L.), cherry guava (P. littorale Raddi), mulberry (Morus nigra L.), loquat (Eriobotrya japonica (Thunb.) Lindl.), and pear (Pyrus communis L.). Therefore, the major citrus crops commercially cultivated in Australia have a relatively low susceptibility to B. tryoni, with Eureka lemons being a particularly poor host for this tephritid fruit fly.

Effectiveness of Various Artificial Larval Diets for Rearing Bactrocera passiflorae (Froggatt) and B. xanthodes (Broun) in the Laboratory in Fiji

Laboratory colonies of Bactrocera passiflorae (Froggatt) and B. xanthodes (Broun) were established at Koronivia Research Station, Fiji in 1991. Laboratory rearing of the two economically important species was a prerequisite to studies conducted on protein bait spray and quarantine treatment development. To increase the production of laboratory reared fruit flies for this research and also to have a substitute larval diet available, replicated comparisons of the effectiveness of larval diets were carried out using B. passiflorae and B. xanthodes. The diets compared were pawpaw/bagasse, dehydrated carrot and diets used for culturing Mediterranean fruit fly (Ceratitis capitata Wiedemann), Oriental fruit fly (B. dorsalis Hendel), melon fly (B. cucurbitae Coquillett) and B. latifrons (Hendel), pawpaw diet and breadfruit diet. B. passiflorae and B. xanthodes eggs seeded onto the various diets were allowed to develop into larvae, pupae and adults. The percentage egg hatch, number of pupae recovered, percentage pupal mortality, weight of 100 pupae, number of adults and percentage eclosion were used to determine the effectiveness of the diets. Results showed that pawpaw/bagasse and dehydrated carrot diets performed favorably for both species. The pawpaw diet currently used as standard larval diets for both species is the most readily available and easiest to use. Breadfruit diet was tested on B. xanthodes only and showed that it was a suitable substitute for the pawpaw-based diets. Other larval diets, cassava/pawpaw and banana diets, that have been developed and used in the South Pacific areas are also discussed in this paper. When pawpaw or breadfruit are not available, dehydrated carrot diet may be substituted for fruit-based larval diets.