Dissertations / Theses on the topic 'Bactrocera tryoni'

This thesis examined the feeding behaviour of B. tryoni relative to the physiological state (sex, maturity, mated state and nutritional condition) of the fly. I particularly focused on the attraction of B. tryoni to proteinaceous food types with the intention of explaining fly response as a function of physiological state. All B. tryoni, irrespective of age, sex or mated state, preferentially feed in the first few hours after sunrise, both in the laboratory and field cage environment. Sugar feeding was greatest by mature mated females, followed closely by immature flies (both sexes). Protein feeding was greatest in immature flies, while mature unmated females spent very little time in protein feeding. Thus, proteinaceous baits used for B. tryoni control would be least effective over mature unmated females. Fly hunger is also relevant in determining how a fly will respond to food types. Some results with A. ludens suggested that sugar hungry flies had a reduced attraction to proteinaceous odours, a relevant consequence that significantly reduces the success of proteinaceous baits. Indeed, gravid female B. tryoni had a much decreased attraction to bacteria when sugar hungry, but the effect was not significant for immature flies (both sexes). Therefore, I tested B. tryoni attraction to sugar food (open fruit) and protein food (bacteria) combined, but the combined odours did not appear to be significantly more attractive than one food source alone. Gravid females were primarily attracted to fruit odour as an oviposition resource, independent of their nutritional condition. Further, gravid females were only significantly attracted bacterial odour when protein hungry and were repelled when fed. These results then lead me to a further examination of fly attraction to bacterial odours. Common phyllosphere bacteria are a known protein source for B. tryoni. Anecdotal evidence suggests that fly attraction to bacteria increases as the bacterial culture ages. However this was only found true for immature flies. Gravid females that were protein hungry initially had a strong response to bacterial odours in the exponential growth phase, but had a much reduced response to stronger bacterial odours in the stationary / death phase. This supports the theory that bacterial odours represent an ovipositional deterrent to B. tryoni. A common volatile released by phyllosphere bacteria, 2-butanone, is also thought to be the attractive volatile in cue lure (a male B. tryoni sex attractant). However, mature males had a relatively low response to bacteria, suggesting that bacterial odours do not act as a sex attractant in the same way as cue lure. My findings help explain why protein baits may or may not be effective in controlling B. tryoni and will improve the decision making process when considering how best to control B. tryoni.
Thesis (Masters)
Master of Philosophy (MPhil)
Griffith School of Environment
Science, Environment, Engineering and Technology
Full Text

Doctor of Philosophy
A number of types of molecular polymorphisms can be used for studying genetic relationship and evolutionary history. Microsatellites are hypervariable and can be very useful tools to determine population structure, distinguish sibling species, as well as verifying parental relationships and pedigrees. However, while microsatellite polymorphisms are useful for solving relationships between populations within a species, relations among species or genera will probably be obscured due to a high degree of homoplasy —identity arising from evolutionary convergence not by descent. For long range evolutionary history, such as phylogeny from old world monkey to human, mtDNA markers may be better candidates. The aim of this thesis is to assess molecular polymorphisms of different types and their optimal use in different situations. Two widely separated taxa were used for testing –the green monkey Chlorocebus sabaeus, and the sibling dipteran flies Bactrocera tryoni and B. neohumeralis, known collectively as the Queensland fruit fly. In the present study a complete 16,550 bp mtDNA sequence of the green monkey Chlorocebus sabaeus is reported for the fist time and has been annotated (Chapter 2). Knowledge of the mtDNA genome contributes not only to identification of large scale single nucleotide polymorphisms (SNPs) (Chapter 4) or other mtDNA polymorphisms development, but also to primate phylogenetic and evolutionary study (Chapter 3). Microsatellites used for the green monkey paternity and pedigree studies were developed by cross-amplification using human primers (Chapter 5). For studies of population structure and species discrimination in Queensland fruit fly (Chapter 7), microsatellites were isolated from a genomic library of Bactrocera tryoni (Chapter 6) The total length of 16550 bp of complete mtDNA of the green monkey C. sabaeus, which has been sequenced and annotated here, adds a new node to the primate phylogenetic tree, and creates great opportunity for SNP marker development. The heteroplasmic region was cloned and five different sequences from a single individual were obtained; the implication of this are discussed. The phylogenetic tree reconstructed using the complete mtDNA sequence of C. sabaeus and other primates was used to solve controversial taxonomic status of C. sabaeus. Phylogenies of primate evolution using different genes from mtDNA are discussed. Primate evolutionary trees using different substitution types are compared and the phylogenetic trees constructed using transversions for the complete mtDNA were found close to preconceived expectations than those with transversions + transitions. The sequence of C. sabaeus 12SrRNA reported here agrees with the one published by ven der Kuyl et al. (1996), but additional SNPs were identified. SNPs for other regions of mtDNA were explored using dHPLC. Twenty two PCR segments for 96 individuals were tested by dHPLC. Fifty five SNPs were found and 10 haplogroups were established. Microsatellite markers were used to construct a genealogy for a colony of green monkeys (C. sabaeus) in the UCLA Vervet Monkey Research Colony. Sixteen microsatellites cross-amplified from human primers were used to conduct paternity analysis and pedigree construction. Seventy-eight out of 417 offspring were assigned paternity successfully. The low success rate is attributed to a certain proportion of mismatches between mothers and offspring; the fact that not all candidate fathers were sampled, the limitations of microsatellite polymorphisms; and weakness of the exclusion method for paternity assessment. Due to the low success rate, the pedigree is split into a few small ones. In a complicated pedigree composed of 75 animals and up to four generations with multiple links a power male mated with 8 females and contributed 10 offspring to the pedigree. Close inbreeding was avoided. Population structure within two species of Queensland fruit fly Bactrocera tryoni and Bactrocera neohumeralis (Tephritidae: Diptera) is examined using microsatellite polymorphisms. Queensland fruit flies B. tryoni and B. neohumeralis are sympatric sibling species that have similar morphological and ecological features. They even share polymorphism at the molecular level. Mating time difference is the main mechanism by which they maintain separate species. In the present study, 22 polymorphic and scorable microsatellites were isolated from B. tryoni and tested in the two species sampled from sympatric distribution areas. Pairwise genetic distance analysis showed explicit differentiation in allele frequencies between the two species, but very weak differences between conspecific populations. Gene flow is higher within B. tryoni than within B. neohumeralis, and gene exchange between the two species exists. An averaging linkage clustering tree constructed by UPGMA showed two major clusters distinguishing the two species, and it appears that population structure is highly correlated with geographic distance. The relationship between molecular markers, evolution, and selection are discussed using comparative studies within two large taxa: primate and insect. The degree of conservation and polymorphism in microsatellites varies between taxa, over evolutionary time.

A number of types of molecular polymorphisms can be used for studying genetic relationship and evolutionary history. Microsatellites are hypervariable and can be very useful tools to determine population structure, distinguish sibling species, as well as verifying parental relationships and pedigrees. However, while microsatellite polymorphisms are useful for solving relationships between populations within a species, relations among species or genera will probably be obscured due to a high degree of homoplasy —identity arising from evolutionary convergence not by descent. For long range evolutionary history, such as phylogeny from old world monkey to human, mtDNA markers may be better candidates. The aim of this thesis is to assess molecular polymorphisms of different types and their optimal use in different situations. Two widely separated taxa were used for testing –the green monkey Chlorocebus sabaeus, and the sibling dipteran flies Bactrocera tryoni and B. neohumeralis, known collectively as the Queensland fruit fly. In the present study a complete 16,550 bp mtDNA sequence of the green monkey Chlorocebus sabaeus is reported for the fist time and has been annotated (Chapter 2). Knowledge of the mtDNA genome contributes not only to identification of large scale single nucleotide polymorphisms (SNPs) (Chapter 4) or other mtDNA polymorphisms development, but also to primate phylogenetic and evolutionary study (Chapter 3). Microsatellites used for the green monkey paternity and pedigree studies were developed by cross-amplification using human primers (Chapter 5). For studies of population structure and species discrimination in Queensland fruit fly (Chapter 7), microsatellites were isolated from a genomic library of Bactrocera tryoni (Chapter 6) The total length of 16550 bp of complete mtDNA of the green monkey C. sabaeus, which has been sequenced and annotated here, adds a new node to the primate phylogenetic tree, and creates great opportunity for SNP marker development. The heteroplasmic region was cloned and five different sequences from a single individual were obtained; the implication of this are discussed. The phylogenetic tree reconstructed using the complete mtDNA sequence of C. sabaeus and other primates was used to solve controversial taxonomic status of C. sabaeus. Phylogenies of primate evolution using different genes from mtDNA are discussed. Primate evolutionary trees using different substitution types are compared and the phylogenetic trees constructed using transversions for the complete mtDNA were found close to preconceived expectations than those with transversions + transitions. The sequence of C. sabaeus 12SrRNA reported here agrees with the one published by ven der Kuyl et al. (1996), but additional SNPs were identified. SNPs for other regions of mtDNA were explored using dHPLC. Twenty two PCR segments for 96 individuals were tested by dHPLC. Fifty five SNPs were found and 10 haplogroups were established. Microsatellite markers were used to construct a genealogy for a colony of green monkeys (C. sabaeus) in the UCLA Vervet Monkey Research Colony. Sixteen microsatellites cross-amplified from human primers were used to conduct paternity analysis and pedigree construction. Seventy-eight out of 417 offspring were assigned paternity successfully. The low success rate is attributed to a certain proportion of mismatches between mothers and offspring; the fact that not all candidate fathers were sampled, the limitations of microsatellite polymorphisms; and weakness of the exclusion method for paternity assessment. Due to the low success rate, the pedigree is split into a few small ones. In a complicated pedigree composed of 75 animals and up to four generations with multiple links a power male mated with 8 females and contributed 10 offspring to the pedigree. Close inbreeding was avoided. Population structure within two species of Queensland fruit fly Bactrocera tryoni and Bactrocera neohumeralis (Tephritidae: Diptera) is examined using microsatellite polymorphisms. Queensland fruit flies B. tryoni and B. neohumeralis are sympatric sibling species that have similar morphological and ecological features. They even share polymorphism at the molecular level. Mating time difference is the main mechanism by which they maintain separate species. In the present study, 22 polymorphic and scorable microsatellites were isolated from B. tryoni and tested in the two species sampled from sympatric distribution areas. Pairwise genetic distance analysis showed explicit differentiation in allele frequencies between the two species, but very weak differences between conspecific populations. Gene flow is higher within B. tryoni than within B. neohumeralis, and gene exchange between the two species exists. An averaging linkage clustering tree constructed by UPGMA showed two major clusters distinguishing the two species, and it appears that population structure is highly correlated with geographic distance. The relationship between molecular markers, evolution, and selection are discussed using comparative studies within two large taxa: primate and insect. The degree of conservation and polymorphism in microsatellites varies between taxa, over evolutionary time.

Queensland fruit fly is a destructive horticultural insect pest. Knowing the age-structure of fly populations, that is the relative proportion of young, middle-age, and old-age flies within a population at a given time, is critical for effective management. The thesis combined behavioural ecology with a novel mathematical analysis to identify the seasonal changes in the age of a wild Queensland fruit fly population. The study showed that the abundance and age-structure of the fly changed predictably with the season, strongly suggestive of an endogenous mechanism that helps the fly cope with seasonal changes in resource availability.

This project elucidated functional role of phytochemicals used in the management of pest fruit flies. Comparative behavioural, physiological and genomic approaches revealed that phytochemicals are mediating reproductive fitness by changing pheromonal compound males release and by making them physiologically more active. The possible mechanistic functions are that the phytochemicals act as a pheromone booster and as an energy supplement.

The research outlined in this thesis primarily focused on the isolation and characterization of representatives of the mariner family of transposable elements in the genome of the tephritid, Bactrocera tryoni (Queensland fruit fly). A preliminary investigation was also made of the mobility properties of constructs based on the mariner element Mos], following their transient introduction into the embryonic soma of B. tryoni. This involved the use of plasmid-based excision and transposition assays. These studies were partly undertaken to obtain an initial idea of the feasibility of developing a germline transformation system based on particular mariner elements for use with B. tryoni, and possibly other related tephritids. To investigate whether the B. tryoni genome contained endogenous mariner element copies, an initial PCR analysis was carried out using degenerate oligonucleotide primers that had previously been designed to stretches of conserved residues within transposases encoded by particular mariner elements. Using this approach, examples of at least five distinct types of mariner elements were detected in the genome of B. tryoni following the cloning and DNA sequencing of 20 unique amplified fragments. Phylogenetic analyses using the conceptual amino acid translations of these partial transposase gene regions indicated that the B. tryom‘ sequences represented three subfamilies of mariner elements. Using the same PCR-based approach, diverse mariner sequences were also isolated from Bactrocera neohumeralis, a sibling species of B. tryoni, and an additional tephritid, Bactrocera jarvisi.

The host use patterns of tephritids (fruit flies) range from those that are highly specific (monophagous, oligophagous) to those that use a wide range of hosts from different plant families (polyphagous). Knowledge of the mechanisms responsible for such host use patterns is critical in understanding the insect-plant relationship and developing successful population management strategies. Several studies have revealed that the behavioural and environmental factors play a significant role in host use. A key theory thought to explain host use patterns of phytophagous insects has been optimality theory (preference-performance hypothesis), however, this has seldom been investigated in fruit flies. In this thesis I compared the host use pattern of the specialist (oligophagous) cucumber fruit fly, Bactrocera cucumis (French), and the generalist (polyphagous) Queensland fruit fly, Bactrocera tryoni (Froggatt) (Diptera: Tephritidae: Dacinae), both serious agricultural pests in Queensland, Australia. I primarily focussed on investigating the role of the preference-performance hypothesis in explaining the host use pattern of these two species at different plant taxonomic levels. I also examined the life history consequences of host use patterns by investigating the demographies of these two dacines on different host plants. The preference-performance hypothesis postulates that the host preference executed by an insect should be positively correlated to its offspring performance. While there were some correlations between preference and performance parameters for both B. cucumis and B. tryoni, these were dependent on the parameter used and were more relevant in explaining host use patterns in B. cucumis than B. tryoni. Both species exhibited significant host discrimination among the different hosts presented to them. More significantly, they preferentially used certain host plants even when hosts of purportedly equivalent status were presented to them simultaneously. The efficiency with which they used their hosts differed even when offered different varieties of a host species from their primary host family. These patterns, observed in both these dacine species, indicated that host use is more plausibly explained by behavioural and physiological factors associated with host use. Key factors affecting host acceptance behaviour and subsequent host use of dacine fruit flies include pre-alighting factors (e.g., host plant structure, fruit colour, shape and size) and post-alighting factors (e.g., pericarp toughness, fruit volatiles and chemical composition of fruit). I investigated the role of fruit size and pericarp toughness in explaining host use patterns. Pericarp toughness appeared to strongly affect host acceptance and subsequent host use in both dacine species. Although some fruit species were readily accepted for oviposition by receiving a high proportion of oviposition attempts from both fly species, very few of these oviposition attempts resulted in pericarp penetration (oviposition event). This behaviour was primarily attributed to the differences in pericarp toughness; fruits with a tougher pericarp provided greater resistance to ovipositor penetration, thus resulting in less use than those with a softer pericarp. In addition to the behavioural aspects of fruit fly-host plant relationships, life history parameters of tephritid species play a significant role in host use. Investigations of the demography of these species on different hosts revealed that the polyphagous B. tryoni has higher fecundity and net reproductive rate, and shorter generation time while the oligophagous B. cucumis has a lower fecundity and net reproductive rate with a longer generation time. While the demographic patterns of the B. cucumis appeared to conform to preference-performance hypothesis, B. tryoni showed some interesting departures from the predictions of the preference-performance hypothesis. For B. tryoni, it was evident that some hosts that may yield lower survival in terms of larval survival (e.g., plum) may still be demographically suitable because of the enhanced reproductive performance of adults emerging from these fruits. This highlights the significance of the quality of adult feeding resource in the demography of this species. Organizing demographic data and analysing them using population projection models also enabled me to identify critical life stages that influence the demography of these two species across different host fruits. This study found that for both the specialist B. cucumis and the generalist B. tryoni, population growth rate is highly sensitive at the adult reproductive stage, indicating that manipulating probability of survival at this life stage would be critical to manage the population of these pest species. This may explain the success of protein bait sprays as a technique to manage these pest species. Predators or pathogens that target the adult reproductive stage may also provide successful biological control of these pest fruit fly populations. In addition to survival of reproductive individuals, pupal survival was also important to the demography of these species, suggesting that biological control targeting pupal stage may also yield population suppression across different fruit production systems. In summary, my studies showed that, while the preference-performance hypothesis may be a useful heuristic tool in explaining comparative host use patterns in the Dacinae, it may be more valuable in understanding primary adaptations of different species that enable them to use different host plants. This understanding combined with the life-history consequences of host use would not only shed light on their ecology, but also prove valuable in designing suitable management strategies for pest dacines.
Thesis (PhD Doctorate)
Doctor of Philosophy (PhD)
Australian School of Environmental Studies
Full Text

Bactrocera tryoni, the Queensland fruit fly, is established along the entire Australian east coast. It is a major pest of horticulture and arguably the worst horticultural insect pest in Australia. Adult flies lay eggs into fruit and resultant larvae feed on the flesh of the fruit. The population biology of B. tryoni has been well studied in temperate regions, where it has been established that climatic factors, particularly temperature and rainfall, limit population growth. In contrast, in subtropical and tropical regions, the population dynamics of the fly have been little studied. This thesis investigates the fly's phenology and abundance changes across subtropical and tropical Queensland and asks what factors govern the population cycles of B. tryoni in this state. Winter breeding and abundance of the fly, a component of the seasonal cycle which in south-east Queensland is fundamentally different from that observed in temperate Australia, is also investigated. A historical, extensive multi-year and multi-site trapping data set with from across Queensland was analysed to look at the effects of temperature, rainfall and relative humidity on B. tryoni trap catch. Trap data was further compared with the predicted phenology data generated by a DYMEX® based B. tryoni population phenology model. The phenology model used was based on a previously published model, but was also modified to more explicitly look at the effects of host plant availability and the presence or absence of non-reproductive over-wintering flies. Over-wintering field cage studies and a winter-spring field trapping study, both carried out in Brisbane, supplied additional data on B. tryoni's population abundance and capacity to breed during winter in the subtropics. Results show significant variation of monthly fly abundance for nine sites across Queensland. Abundance changed across sites in non-predictable ways. Annual population phenology within a site was, for some sites, highly consistent from year to year, but inconsistent for other sites. All sites in the subtropics showed some form of population depression during the cooler months, but breeding was continuous, albeit reduced at nearly all sites. Some tropical sites, where the climate is regarded as highly favourable for B. tryoni, still showed dramatic peaks and troughs in annual population abundance. There were relatively few significant correlations observed between weather factors and fly populations for any site. Output from the DYMEX population model suggested that fruit availability is a major driver of population dynamics in the tropical north of the state, while weather is more important in the subtropical south. The population dynamics of B. tryoni at sites along the central Queensland coast, where it is assumed that a mix of both weather and host fruit availability drive local populations, were poorly captured by the population model. Field cage results showed that B. tryoni successfully bred during winter in Brisbane, with pupal emergence starting in mid-winter (1st week of August), peaking in early spring (2nd week of September). Trap catch at orchards in Brisbane increased with increasing temperature and fruit availability, but diminished with decreasing temperature and fruit availability. The results suggest that B. tryoni has an optimal climate for population growth in the tropics, but fruit availability for offspring production limits population growth. In the subtropics however, both climate and fruit availability determine the population size. Winter temperatures are marginal for B. tryoni population growth in the subtropics.

Bactrocera tryoni, the Queensland fruit fly, is established along the entire Australian east coast. It is a major pest of horticulture and arguably the worst horticultural insect pest in Australia. Adult flies lay eggs into fruit and resultant larvae feed on the flesh of the fruit. The population biology of B. tryoni has been well studied in temperate regions, where it has been established that climatic factors, particularly temperature and rainfall, limit population growth. In contrast, in subtropical and tropical regions, the population dynamics of the fly have been little studied. This thesis investigates the fly's phenology and abundance changes across subtropical and tropical Queensland and asks what factors govern the population cycles of B. tryoni in this state. Winter breeding and abundance of the fly, a component of the seasonal cycle which in south-east Queensland is fundamentally different from that observed in temperate Australia, is also investigated. A historical, extensive multi-year and multi-site trapping data set with from across Queensland was analysed to look at the effects of temperature, rainfall and relative humidity on B. tryoni trap catch. Trap data was further compared with the predicted phenology data generated by a DYMEX® based B. tryoni population phenology model. The phenology model used was based on a previously published model, but was also modified to more explicitly look at the effects of host plant availability and the presence or absence of non-reproductive over-wintering flies. Over-wintering field cage studies and a winter-spring field trapping study, both carried out in Brisbane, supplied additional data on B. tryoni's population abundance and capacity to breed during winter in the subtropics. Results show significant variation of monthly fly abundance for nine sites across Queensland. Abundance changed across sites in non-predictable ways. Annual population phenology within a site was, for some sites, highly consistent from year to year, but inconsistent for other sites. All sites in the subtropics showed some form of population depression during the cooler months, but breeding was continuous, albeit reduced at nearly all sites. Some tropical sites, where the climate is regarded as highly favourable for B. tryoni, still showed dramatic peaks and troughs in annual population abundance. There were relatively few significant correlations observed between weather factors and fly populations for any site. Output from the DYMEX population model suggested that fruit availability is a major driver of population dynamics in the tropical north of the state, while weather is more important in the subtropical south. The population dynamics of B. tryoni at sites along the central Queensland coast, where it is assumed that a mix of both weather and host fruit availability drive local populations, were poorly captured by the population model. Field cage results showed that B. tryoni successfully bred during winter in Brisbane, with pupal emergence starting in mid-winter (1st week of August), peaking in early spring (2nd week of September). Trap catch at orchards in Brisbane increased with increasing temperature and fruit availability, but diminished with decreasing temperature and fruit availability. The results suggest that B. tryoni has an optimal climate for population growth in the tropics, but fruit availability for offspring production limits population growth. In the subtropics however, both climate and fruit availability determine the population size. Winter temperatures are marginal for B. tryoni population growth in the subtropics.

Fruit flies are serious pests of fruit around the world, as their maggots destroy fruit by feeding within them. An important non-pesticide control option for fruit flies is the use of natural enemies. This study investigated the role of memory and learning in the host searching behaviour of one such natural enemy, the small wasp, Diachasmimorpha kraussii. This wasp is native to Australia and attacks our most serious fruit fly pest, the Queensland Fruit Fly. This research studied fundamental insect behaviour, but produced specific recommendations for the more effective use of Diachasmimorpha kraussii for sustainable pest management.

Queensland fruit fly is Australia’s most destructive horticultural insect pest. The flies need to mate to successfully reproduce, but there remained significant gaps in knowledge about how they find and select mates. I showed that male and female flies likely use physical landmarks to find each other in the environment. Having found potential mates, I described their fine-scale courtship behaviour and demonstrated that young, large male flies are most successful at securing a mate. I also made significant advances in our understanding of the potential for close-range chemical communication to play a role in mate identification and selection. This research directly informs sustainable management strategies against this pest.

Fruit flies are the insects which cause maggots in your backyard fruit and vegetables. They are not just a nuisance to gardeners, but the single greatest insect threat to commercial and subsistence fruit growers throughout Asia, Australia and the Pacific. Queensland fruit fly, the focus of this PhD, costs Australia an estimated $100million per year. I focused specifically on how Queensland fruit fly uses different commercial citrus varieties. I identified specific plant related mechanisms which increase a fruit’s resistance to fruit fly attack. This information can be used by plant breeders to make fruit less prone to fruit fly damage.

Outbreaks of insect pests pose a serious threat to local economies and global food production, with as much as 15% of global crop production lost to herbivorous insects annually. Outbreaks of transboundary pests and diseases that affect food crops have increased in frequency in conjunction with globalization, international trade and the impacts of climate change. Indeed, increasing global temperatures are predicted to increase the distribution, rate of development, survival and population density of many pest insects. Such changes have important ramifications for host plant exploitation. The Queensland fruit fly (Bactrocera tryoni) is Australia’s worst horticultural pest, and is feared by international buyers of Australian produce. Like other Tephritid fruit fly species, B. tryoni has the potential to breach quarantine barriers via human mediated transport, and can rapidly establish in ‘new’ environments. This pest species is responsible for an estimated AU$28.5 million in annual yield loss, management costs and loss to domestic and international markets. Increasing and ongoing outbreaks of B. tryoni in Australia’s major growing regions has put international trade in jeopardy.
Thesis (PhD Doctorate)
Doctor of Philosophy (PhD)
Griffith School of Environment
Science, Environment, Engineering and Technology
Full Text

The Queensland fruit fly, Bactrocera tryoni (Froggatt) (Diptera: Tephritidae), a major pest of horticulture in eastern Australia, is a relatively poor coloniser of new habitat. This thesis examines behavioural properties that might limit the ability of B. tryoni to establish new populations. As the potential for B. tryoni to establish an outbreak population may be most directly limited by mechanisms associated with dispersal and mating behaviour, these two factors were the focus of this research project. The relevance of dispersal and mating behaviour for control of outbreak populations was assessed. Dispersal (i) Dispersal patterns of males and females are not different. Dispersal of post-teneral male B. tryoni from a point within an orchard near Richmond, New South Wales, was monitored following temporally replicated releases. Application of sterile insect technique (SIT) requires knowledge of dispersal from a release point so that effective release rates can be determined. In addition, dispersal following introduction to new habitat can lead to low or negative population growth and an Allee effect. In Spring and Autumn, 2001 – 2003, three different strains of B. tryoni were released: (1) wild flies reared from infested fruit collected in the Sydney Basin; (2) a laboratory-reared strain with a colour mutation (white marks); and (3) sterile flies obtained by gamma-irradiation of a mass-reared strain. Dispersal was monitored using a grid of traps baited with the male attractant, cuelure. During the majority of releases, flies were massmarked using a self-marking technique and fluorescent pigment powder to enable identification of recaptured flies. A preliminary study found that fluorescent pigment marks had no effect on adult survival and marks did not fade significantly in the laboratory over a period of five weeks after eclosion. As cuelure repels inseminated sexually mature female B. tryoni, unbaited, coloured flat sticky traps, and black and yellow sticky sphere traps baited with a food lure (protein autolysate solution) were used to supplement traps baited with cuelure. The effectiveness of these two sticky trap types was assessed, and recaptures used to compare patterns of dispersal from a release point by male and female B. tryoni. Fluorescent yellow (chartreuse), green, and clear unbaited flat sticky traps were relatively ineffective for monitoring dispersal of sterile male and female B. tryoni, recapturing only 0.1% of released sterile flies. Monitoring dispersal with sticky ball traps baited with protein autolysate solution was more successful, with yellow spheres and black spheres recapturing 1.7% and 1.5%, respectively. Trap colour had no effect on recaptures on flat sticky traps or sticky spheres. Equal recapture rates on yellow and black sticky sphere traps suggests that the odour of yeast autolysate solution was more important than colour for attraction of post-teneral flies to traps. Using the results of recaptures on odoriferous black and yellow sticky sphere traps within one week of release, regression equations of male and female recaptures per trap were found to be similar (Figure 4-3). This is the first study to clearly indicate that post-teneral dispersal patterns of male and female B. tryoni released from a point do not differ, enabling the use of existing models to predict density of both sexes of B. tryoni following post-teneral dispersal. (ii) Males disperse further in Spring than in Autumn, but this is not temperature-related. Analysis of replicated recaptures in traps baited with cuelure revealed that dispersal of male B. tryoni in an orchard near Richmond, New South Wales, was higher in Spring than in Autumn (Figure 5-6). As the maximum daily temperature was significantly higher in Spring than in Autumn this result was unexpected, since earlier studies have found that B. tryoni disperse at the onset of cool weather in search of sheltered over-wintering sites. Dispersal of post-teneral B. tryoni may have been affected by habitat suitability; it was found that seasonal trends in dispersal could have been influenced by local habitat variables. Low mean dispersal distances in Autumn may be explained by the presence of fruiting hosts in the orchard, or the availability of resources required by over-wintering flies. There was no significant correlation between temperature and mean dispersal distance, suggesting that higher rates of dispersal cannot be explained by temperature-related increases in activity. Recapture rate per trap was significantly negatively correlated with increasing daily maximum and average temperature. This may have consequences for detection of B. tryoni outbreaks in quarantine areas due to reduced cuelure trap efficiency. (iii) Maturity and source variation affect dispersal and response to cuelure. This research indicated that most male and female B. tryoni do not disperse far from a release point, suggesting that an invading propagule would not spread far in the first generation. However, there is considerable variation in flight capability among individuals. Comparison of wild, laboratory-reared white marks, and gamma-irradiated sterile male B. tryoni indicated that mean dispersal distance and redistribution patterns were not significantly affected by fly origin. Despite no difference in dispersal distance from the release point, recaptures of wild and sterile males per Lynfield trap baited with cuelure were highest within one week after release, whereas recaptures of white marks males per trap increased in the second week. This result may offer evidence to support the hypothesis that sterile male B. tryoni respond to cuelure at an earlier age. Rearing conditions used to produce large quantities of males for sterilisation by gamma-irradiation may select for earlier sexual maturity. Mating Behaviour (i) Density and sex ratio do not affect mating, except at low densities. Demographic stochasticity in the form of sex ratio fluctuations at low population density can lead to an observed Allee effect. The effect of local group density and sex ratio on mating behaviour and male mating success of a laboratory-adapted strain of B. tryoni was examined in laboratory cages. In the laboratory-adapted strain of B. tryoni used in this study, a group of one female and one male was sufficient for a good chance of mating success. The proportion of females mated and male mating success was not significantly affected by density or sex ratio, although variability in male mating success was higher at low density. This could indicate that mating success of B. tryoni can be reduced when local group density is low owing to decreased frequency in encounters between males and females. (ii) Mass-reared males exhibit aberrant mating behaviour, but this does not reduce mating success. Strong artificial selection in mass-rearing facilities may lead to decreased competitiveness of sterile males released in SIT programs as a result of alteration or loss of ecological and behavioural traits required in the field. The effects of domestication and irradiation on the mating behaviour of males of B. tryoni were investigated by caging wild, mass-reared and sterile (mass-reared and gammairradiated) males with wild females. Mating behaviour of mass-reared males was different from that of wild males, but behaviour of wild and sterile males was similar. Mass-reared males were found to engage in mounting of other males much more frequently than wild and sterile males, and began calling significantly earlier before darkness. Male calling did not appear to be associated with female choice of mating partners, although this does not exclude the possibility that calling is a cue used by females to discriminate between mating partners. Conditions used to domesticate and rear large quantities of B. tryoni for SIT may select for an alternative male mating strategy, with mass-reared males calling earlier and exercising less discrimination between potential mating partners. Despite differences in behaviour of wild, mass-reared and sterile males, frequency of successful copulations and mating success were similar. (iii) Pheromone-calling by males was increased in larger aggregations but this did not result in significantly more female visits. Finally, large laboratory cages with artificial leks were used to investigate the importance in B. tryoni of male group size for female visitation at lek sites and initiation of male pheromone-calling. Calling propensity of male B. tryoni was increased by the presence of conspecific males. Females visited the largest lek more frequently than single males, but there was no correlation between lek size and female visitation. Female B. tryoni had a limited capacity to perceive a difference between the number of calling males; female visitation at leks was only weakly associated with male calling, suggesting that lek size and the number of pheromone-calling males may not be the only factor important in locating mates in B. tryoni. The weak, but positive correlation between male calling and female visitation may indicate that passive attraction maintains lek-mating in B. tryoni. Further studies are essential on mating behaviour of B. tryoni, including identification of male mating aggregations in the field, measurement of habitat variables associated with male aggregations, the influence of density on wild B. tryoni mating success, and the role of pheromone-calling, in order to optimise use of SIT for control of this pest.

The Queensland fruit fly, Bactrocera tryoni (Froggatt) (Diptera: Tephritidae), a major pest of horticulture in eastern Australia, is a relatively poor coloniser of new habitat. This thesis examines behavioural properties that might limit the ability of B. tryoni to establish new populations. As the potential for B. tryoni to establish an outbreak population may be most directly limited by mechanisms associated with dispersal and mating behaviour, these two factors were the focus of this research project. The relevance of dispersal and mating behaviour for control of outbreak populations was assessed. Dispersal (i) Dispersal patterns of males and females are not different. Dispersal of post-teneral male B. tryoni from a point within an orchard near Richmond, New South Wales, was monitored following temporally replicated releases. Application of sterile insect technique (SIT) requires knowledge of dispersal from a release point so that effective release rates can be determined. In addition, dispersal following introduction to new habitat can lead to low or negative population growth and an Allee effect. In Spring and Autumn, 2001 � 2003, three different strains of B. tryoni were released: (1) wild flies reared from infested fruit collected in the Sydney Basin; (2) a laboratory-reared strain with a colour mutation (white marks); and (3) sterile flies obtained by gamma-irradiation of a mass-reared strain. Dispersal was monitored using a grid of traps baited with the male attractant, cuelure. During the majority of releases, flies were massmarked using a self-marking technique and fluorescent pigment powder to enable identification of recaptured flies. A preliminary study found that fluorescent pigment marks had no effect on adult survival and marks did not fade significantly in the laboratory over a period of five weeks after eclosion. As cuelure repels inseminated sexually mature female B. tryoni, unbaited, coloured flat sticky traps, and black and yellow sticky sphere traps baited with a food lure (protein autolysate solution) were used to supplement traps baited with cuelure. The effectiveness of these two sticky trap types was assessed, and recaptures used to compare patterns of dispersal from a release point by male and female B. tryoni. Fluorescent yellow (chartreuse), green, and clear unbaited flat sticky traps were relatively ineffective for monitoring dispersal of sterile male and female B. tryoni, recapturing only 0.1% of released sterile flies. Monitoring dispersal with sticky ball traps baited with protein autolysate solution was more successful, with yellow spheres and black spheres recapturing 1.7% and 1.5%, respectively. Trap colour had no effect on recaptures on flat sticky traps or sticky spheres. Equal recapture rates on yellow and black sticky sphere traps suggests that the odour of yeast autolysate solution was more important than colour for attraction of post-teneral flies to traps. Using the results of recaptures on odoriferous black and yellow sticky sphere traps within one week of release, regression equations of male and female recaptures per trap were found to be similar (Figure 4-3). This is the first study to clearly indicate that post-teneral dispersal patterns of male and female B. tryoni released from a point do not differ, enabling the use of existing models to predict density of both sexes of B. tryoni following post-teneral dispersal. (ii) Males disperse further in Spring than in Autumn, but this is not temperature-related. Analysis of replicated recaptures in traps baited with cuelure revealed that dispersal of male B. tryoni in an orchard near Richmond, New South Wales, was higher in Spring than in Autumn (Figure 5-6). As the maximum daily temperature was significantly higher in Spring than in Autumn this result was unexpected, since earlier studies have found that B. tryoni disperse at the onset of cool weather in search of sheltered over-wintering sites. Dispersal of post-teneral B. tryoni may have been affected by habitat suitability; it was found that seasonal trends in dispersal could have been influenced by local habitat variables. Low mean dispersal distances in Autumn may be explained by the presence of fruiting hosts in the orchard, or the availability of resources required by over-wintering flies. There was no significant correlation between temperature and mean dispersal distance, suggesting that higher rates of dispersal cannot be explained by temperature-related increases in activity. Recapture rate per trap was significantly negatively correlated with increasing daily maximum and average temperature. This may have consequences for detection of B. tryoni outbreaks in quarantine areas due to reduced cuelure trap efficiency. (iii) Maturity and source variation affect dispersal and response to cuelure. This research indicated that most male and female B. tryoni do not disperse far from a release point, suggesting that an invading propagule would not spread far in the first generation. However, there is considerable variation in flight capability among individuals. Comparison of wild, laboratory-reared white marks, and gamma-irradiated sterile male B. tryoni indicated that mean dispersal distance and redistribution patterns were not significantly affected by fly origin. Despite no difference in dispersal distance from the release point, recaptures of wild and sterile males per Lynfield trap baited with cuelure were highest within one week after release, whereas recaptures of white marks males per trap increased in the second week. This result may offer evidence to support the hypothesis that sterile male B. tryoni respond to cuelure at an earlier age. Rearing conditions used to produce large quantities of males for sterilisation by gamma-irradiation may select for earlier sexual maturity. Mating Behaviour (i) Density and sex ratio do not affect mating, except at low densities. Demographic stochasticity in the form of sex ratio fluctuations at low population density can lead to an observed Allee effect. The effect of local group density and sex ratio on mating behaviour and male mating success of a laboratory-adapted strain of B. tryoni was examined in laboratory cages. In the laboratory-adapted strain of B. tryoni used in this study, a group of one female and one male was sufficient for a good chance of mating success. The proportion of females mated and male mating success was not significantly affected by density or sex ratio, although variability in male mating success was higher at low density. This could indicate that mating success of B. tryoni can be reduced when local group density is low owing to decreased frequency in encounters between males and females. (ii) Mass-reared males exhibit aberrant mating behaviour, but this does not reduce mating success. Strong artificial selection in mass-rearing facilities may lead to decreased competitiveness of sterile males released in SIT programs as a result of alteration or loss of ecological and behavioural traits required in the field. The effects of domestication and irradiation on the mating behaviour of males of B. tryoni were investigated by caging wild, mass-reared and sterile (mass-reared and gammairradiated) males with wild females. Mating behaviour of mass-reared males was different from that of wild males, but behaviour of wild and sterile males was similar. Mass-reared males were found to engage in mounting of other males much more frequently than wild and sterile males, and began calling significantly earlier before darkness. Male calling did not appear to be associated with female choice of mating partners, although this does not exclude the possibility that calling is a cue used by females to discriminate between mating partners. Conditions used to domesticate and rear large quantities of B. tryoni for SIT may select for an alternative male mating strategy, with mass-reared males calling earlier and exercising less discrimination between potential mating partners. Despite differences in behaviour of wild, mass-reared and sterile males, frequency of successful copulations and mating success were similar. (iii) Pheromone-calling by males was increased in larger aggregations but this did not result in significantly more female visits. Finally, large laboratory cages with artificial leks were used to investigate the importance in B. tryoni of male group size for female visitation at lek sites and initiation of male pheromone-calling. Calling propensity of male B. tryoni was increased by the presence of conspecific males. Females visited the largest lek more frequently than single males, but there was no correlation between lek size and female visitation. Female B. tryoni had a limited capacity to perceive a difference between the number of calling males; female visitation at leks was only weakly associated with male calling, suggesting that lek size and the number of pheromone-calling males may not be the only factor important in locating mates in B. tryoni. The weak, but positive correlation between male calling and female visitation may indicate that passive attraction maintains lek-mating in B. tryoni. Further studies are essential on mating behaviour of B. tryoni, including identification of male mating aggregations in the field, measurement of habitat variables associated with male aggregations, the influence of density on wild B. tryoni mating success, and the role of pheromone-calling, in order to optimise use of SIT for control of this pest.

This thesis examines the effects of malathion, chlorpyrifos, fipronil and spinosad mixed in fruit fly protein bait on attraction, feeding and mortality of the Queensland fruit fly, Bactrocera tryoni (Froggatt). The effects of weathering of the protein bait-insecticide mixtures on the mortality of B. tryoni were also measured along with attraction, feeding response and toxicity of the protein bait sprays on important arthropod natural enemies particularly the red scale parasitic wasps, Aphytis lingnanensis (Compere) and Comperiella bifasciata (Howard), the green lace wing, Chrysoperla carnea (Stephens) and the mealy bug predator, Cryptolaemus montrouzieri (Mulsant). In field cage experiments, protein-starved male B. tryoni showed the same level of attraction to protein baits mixed with malathion, chlorpyrifos, fipronil, spinosad and protein alone used as the control. However, protein-starved females elicited a difference in attraction with protein baits containing chlorpyrifos and spinosad. Traps with spinosad bait mixtures captured significantly more females compared to traps containing chlorpyrifos bait mixtures. Laboratory feeding experiments on protein-starved female flies demonstrated that baits containing malathion and chlorpyrifos deterred flies from feeding on them. In contrast, no such deterrence was detected with baits containing spinosad, fipronil and protein alone. These results demonstrated that the type of toxicant mixed with protein bait sprays can influence the attraction and feeding responses of B. tryoni. Therefore the process of screening toxicants for use in protein bait mixtures is important and should entail field and laboratory tests. In terms of toxicity, protein baits mixed with malathion and chlorpyrifos caused significantly higher fly mortality and demonstrated a more rapid fly knock down than did spinosad, fipronil and protein alone as the control. Spinosad however was a slow acting toxicant, causing a gradual increase in fly mortality over time. Fly mortality obtained with protein bait mixtures containing malathion, chlorpyrifos and fipronil, applied on citrus leaves, and weathered out doors for up to 6 days did not vary significantly from freshly applied baits. However, the residual effectiveness of bait mixed with spinosad waned significantly after 3 days of out door weathering. Fly mortality caused by 3 day aged spinosad bait mixture was significantly lower than fresh bait mixtures, suggesting a rapid break down of spinosad under field conditions. The parasitism rates of the two most important parasitoids of red scales, A. lingnanensis and C. bifasciata, measured before and after commencement of fruit fly bait spraying in two commercial citrus orchards, did not show any significant negative trend. Aphytis were not attracted to Pinnacle protein, the most commonly used fruit fly protein lure in Australia. A significantly higher number of Aphytis were attracted to honey solution and protein bait mixed with 20% sugar, compared to protein bait alone. A no-choice test further confirmed this result, demonstrating no difference in attraction between protein and water. Protein bait containing malathion, chlorpyrifos, fipronil, and spinosad, fresh and weathered for up to 12 days, caused high mortality in Aphytis. In contrast protein bait mixed with spinosad caused a lower Aphytis mortality after 12 days out door weathering, compared to chlorpyrifos bait mixture. However, for other weathering periods, no such differences in mortality between the treatments were observed. In addition, the parasitizing capacity of Aphytis, after being exposed to these weathered residues, was reduced. Except for the control, aged bait mixtures significantly reduced the fecundity of Aphytis. Therefore, while integrating chemical based field control along with bio-control agents, appropriate measures should be in place to reduce the negative impacts of toxic residues. Overall, the findings from this study indicate that spinosad is a suitable alternative to the older toxicants, for incorporation into fruit fly protein baits. Moreover, fruit fly protein baits in the field are less likely to disrupt the activites of important natural enemies. However, chemical-based control of insect pests in a cropping system should be designed carefully with the objective to prevent potential harm to susceptible biological control agents such as Aphytis.
Thesis (Masters)
Master of Philosophy (MPhil)
Griffith School of Environment
Faculty of Science, Environment, Engineering and Technology
Full Text

Fruit flies are insect pests which cause devastating losses to fruit and vegetable crops around the globe. Replacing pesticides with natural enemies is a preferred option for their sustainable control. This study investigated the abundance, distribution and biology of two such natural enemies: the small parasitic wasps Fopius arisanus and Diachasmimorpha kraussii. F. arisanus was identified as having more successful characteristics than D. kraussii, making it the most common fruit fly parasitod in southeast Queensland. The research lead to advances in understanding insect behaviour as well as recommendations for better utilisation of parasitoids for fruit fly control.

Queensland fruit fly is Australia's most serious insect pest of horticulture. The fly lays its eggs into fruit, where they hatch into maggots which destroy the fruit. Understanding egg laying behaviour, known as oviposition, is a critical but under-researched aspect of fruit fly biology. This thesis focused on three aspects of oviposition: the role of fruit peel as a physical barrier to oviposition; the quality of fruit for maggot development; and the structure and wear of the egg laying organ – the ovipositor. Results showed that flies selected fruit based on their suitability for offspring survival, not because of the softness or hardness of fruit peel. Previously reported use of holes or wounds in fruit peel by ovipositing females was determined to be a mechanism which saved the female time, not a mechanism to reduce ovipositor wear. The results offer insights into the evolution of host use by fruit flies and their sustainable management.

Fruit flies are major pests to a wide range of fruits and vegetables as their larvae cause damage and yield loss. To replace pesticide-based controls with more sustainable management approaches, we need to develop new generation technologies. This thesis focuses on molecular and whole-organism studies to investigate the interactions between Queensland fruit fly larvae and tomato fruit. Through molecular analysis, different mechanisms were identified to determine the susceptibility of various tomato varieties to fruit fly damage. The results pave the way for future studies on breeding for fruit resistance to fruit fly attack and sustainable pest management.

Frugivorous insects, specialised herbivores that consume fruit and seeds, are considered detrimental to host plant fitness. Their direct link to genetic fitness via consumption of plant reproductive tissue, and their negative socioeconomic association with agriculture exacerbates their harmful status. However, empirical testing of insect frugivore effects on host plants, and ecological research on the contribution of insect frugivores to multitrophic frugivory systems, is lacking. In the current study, direct effects of a non-mutualistic, insect frugivore/host plant system were tested and results showed variable effects. Beneficial, detrimental, but predominantly neutral effects on germination and seed production were observed between the Queensland fruit fly (Bactrocera tryoni) and tomato and capsicum plants. Significant effects on seed production were unexpected because infestation occurs after seed set. It was also found that eggplant, although a recorded host of B. tryoni, is inconsistent in its ability to sustain B. tryoni larvae through to its final instar. These results confirmed a simplification and presumption associated with insect frugivore (specifically fruit fly)/host plant interactions. Larval movement, infestation-induced fruit decay, pulp removal and germination were then investigated. For all hosts (tomato, apple and paw paw), treatments infested by B. tryoni decayed significantly quicker and to a greater extent than uninfested treatments, with obvious but variable changes to the texture and appearance. The movement of B. tryoni larvae, pattern of infestation-induced decay and pulp removal was unique and host dependent for all hosts. Only seeds from infested tomato were shown to germinate during the experiment. This indicated that host fruit characteristics are responsible, in part, for variable direct effects on host plant fitness by insect frugivores. Variable direct effects between insect frugivores and host plants, combined with the more rapid decay of infested fruits is likely to have implications for seed dispersal and seed predation by a third trophic level. The characteristics of fruit that are changed by infestation by an insect frugivore were then tested for their effect on a vertebrate frugivore, to illustrate the importance of recognising multitrophic interactions and indirect effects in frugivory. Specifically, seed predating rodents were incorporated into the study and their response to infested and uninfested fruits were recorded, as well as their reaction to the changes in fruit caused by insect frugivores (i.e. texture, smell, larvae presence and sound). Apple and pear infested with B. tryoni larvae were found to attract rodents, while infested tomato and paw paw had a neutral effect on the native rats. This differed from the predominant finding in the literature, which was a deterrent effect on avian seed dispersers. Vertebrate response to fruit infested with insect frugivores therefore, is variable. Assessing the indirect effect of insect frugivores on host plant fitness by attracting or deterring another trophic level requires knowledge of the direct effect between the introduced trophic level and the host plant. For example, the attraction of a seed predator may be as detrimental to host plant fitness as the deterrence of a seed disperser. This illustrates the complexity associated with assessing insect frugivore effects on host plant fitness. Results also indicated that differences in pulp texture, caused by infestation, have a significant effect on rodent preference for infested or uninfested treatments. Pulp texture is likely to effect rodent foraging efficiency, whereas the presence of B. tryoni larvae was observed to be inconsequential to rodent response to fruits. For rodents, and indeed any trophic level motivated by foraging efficiency, this finding raises the issue that for long lived fruiting plants, outside factors such as food abundance and competition for food, may cause a variable response to fruits infested by insect frugivores. From these investigations it has become apparent that insect frugivores are not consistently harmful to host plant fitness, as suggested by their negative stigma, but are likely to contribute variable effects, directly and indirectly, on multiple components of plant fitness and multitrophic frugivory systems.

Frugivorous insects, specialised herbivores that consume fruit and seeds, are considered detrimental to host plant fitness. Their direct link to genetic fitness via consumption of plant reproductive tissue, and their negative socioeconomic association with agriculture exacerbates their harmful status. However, empirical testing of insect frugivore effects on host plants, and ecological research on the contribution of insect frugivores to multitrophic frugivory systems, is lacking. In the current study, direct effects of a non-mutualistic, insect frugivore/host plant system were tested and results showed variable effects. Beneficial, detrimental, but predominantly neutral effects on germination and seed production were observed between the Queensland fruit fly (Bactrocera tryoni) and tomato and capsicum plants. Significant effects on seed production were unexpected because infestation occurs after seed set. It was also found that eggplant, although a recorded host of B. tryoni, is inconsistent in its ability to sustain B. tryoni larvae through to its final instar. These results confirmed a simplification and presumption associated with insect frugivore (specifically fruit fly)/host plant interactions. Larval movement, infestation-induced fruit decay, pulp removal and germination were then investigated. For all hosts (tomato, apple and paw paw), treatments infested by B. tryoni decayed significantly quicker and to a greater extent than uninfested treatments, with obvious but variable changes to the texture and appearance. The movement of B. tryoni larvae, pattern of infestation-induced decay and pulp removal was unique and host dependent for all hosts. Only seeds from infested tomato were shown to germinate during the experiment. This indicated that host fruit characteristics are responsible, in part, for variable direct effects on host plant fitness by insect frugivores. Variable direct effects between insect frugivores and host plants, combined with the more rapid decay of infested fruits is likely to have implications for seed dispersal and seed predation by a third trophic level. The characteristics of fruit that are changed by infestation by an insect frugivore were then tested for their effect on a vertebrate frugivore, to illustrate the importance of recognising multitrophic interactions and indirect effects in frugivory. Specifically, seed predating rodents were incorporated into the study and their response to infested and uninfested fruits were recorded, as well as their reaction to the changes in fruit caused by insect frugivores (i.e. texture, smell, larvae presence and sound). Apple and pear infested with B. tryoni larvae were found to attract rodents, while infested tomato and paw paw had a neutral effect on the native rats. This differed from the predominant finding in the literature, which was a deterrent effect on avian seed dispersers. Vertebrate response to fruit infested with insect frugivores therefore, is variable. Assessing the indirect effect of insect frugivores on host plant fitness by attracting or deterring another trophic level requires knowledge of the direct effect between the introduced trophic level and the host plant. For example, the attraction of a seed predator may be as detrimental to host plant fitness as the deterrence of a seed disperser. This illustrates the complexity associated with assessing insect frugivore effects on host plant fitness. Results also indicated that differences in pulp texture, caused by infestation, have a significant effect on rodent preference for infested or uninfested treatments. Pulp texture is likely to effect rodent foraging efficiency, whereas the presence of B. tryoni larvae was observed to be inconsequential to rodent response to fruits. For rodents, and indeed any trophic level motivated by foraging efficiency, this finding raises the issue that for long lived fruiting plants, outside factors such as food abundance and competition for food, may cause a variable response to fruits infested by insect frugivores. From these investigations it has become apparent that insect frugivores are not consistently harmful to host plant fitness, as suggested by their negative stigma, but are likely to contribute variable effects, directly and indirectly, on multiple components of plant fitness and multitrophic frugivory systems.

International market access for fresh commodities is regulated by international accepted phytosanitary guidelines, the objectives of which are to reduce the biosecurity risk of plant pest and disease movement. Papua New Guinea (PNG) has identified banana as a potential export crop and to help meet international market access requirements, this thesis provides information for the development of a pest risk analysis (PRA) for PNG banana fruit. The PRA is a three step process which first identifies the pests associated with a particular commodity or pathway, then assesses the risk associated with those pests, and finally identifies risk management options for those pests if required. As the first step of the PRA process, I collated a definitive list on the organisms associated with the banana plant in PNG using formal literature, structured interviews with local experts, grey literature and unpublished file material held in PNG field research stations. I identified 112 organisms (invertebrates, vertebrate, pathogens and weeds) associated with banana in PNG, but only 14 of these were reported as commonly requiring management. For these 14 I present detailed information summaries on their known biology and pest impact. A major finding of the review was that of the 14 identified key pests, some research information occurs for 13. The single exception for which information was found to be lacking was Bactrocera musae (Tryon), the banana fly. The lack of information for this widely reported ‘major pest on PNG bananas’ would hinder the development of a PNG banana fruit PRA. For this reason the remainder of the thesis focused on this organism, particularly with respect to generation of information required by the PRA process. Utilising an existing, but previously unanalysed fruit fly trapping database for PNG, I carried out a Geographic Information System analysis of the distribution and abundance of banana in four major regions of PNG. This information is required for a PRA to determine if banana fruit grown in different parts of the country are at different risks from the fly. Results showed that the fly was widespread in all cropping regions and that temperature and rainfall were not significantly correlated with banana fly abundance. Abundance of the fly was significantly correlated (albeit weakly) with host availability. The same analysis was done with four other PNG pest fruit flies and their responses to the environmental factors differed to banana fly and each other. This implies that subsequent PRA analyses for other PNG fresh commodities will need to investigate the risk of each of these flies independently. To quantify the damage to banana fruit caused by banana fly in PNG, local surveys and one national survey of banana fruit infestation were carried out. Contrary to expectations, infestation was found to be very low, particularly in the widely grown commercial cultivar, Cavendish. Infestation of Cavendish fingers was only 0.41% in a structured, national survey of over 2 700 banana fingers. Follow up laboratory studies showed that fingers of Cavendish, and another commercial variety Lady-finger, are very poor hosts for B. musae, with very low host selection rates by female flies and very poor immature survival. An analysis of a recent (within last decade) incursion of B. musae into the Gazelle Peninsula of East New Britain Province, PNG, provided the final set of B. musae data. Surveys of the fly on the peninsular showed that establishment and spread of the fly in the novel environment was very rapid and thus the fly should be regarded as being of high biosecurity concern, at least in tropical areas. Supporting the earlier impact studies, however, banana fly has not become a significant banana fruit problem on the Gazelle, despite bananas being the primary starch staple of the region. The results of the research chapters are combined in the final Discussion in the form of a B. musae focused PRA for PNG banana fruit. Putting the thesis in a broader context, the Discussion also deals with the apparent discrepancy between high local abundance of banana fly and very low infestation rates. This discussion focuses on host utilisation patterns of specialist herbivores and suggests that local pest abundance, as determined by trapping or monitoring, need not be good surrogate for crop damage, despite this linkage being implicit in a number of international phytosanitary protocols.

The digestive tracts of pest fruit flies (Tephritidae: Diptera) contain a diverse range of bacteria. Since the 1980’s there has been increasing interest in the role of microbial symbionts in tephritid fruit fly performance, rising sharply in the past decade. The sterile insect technique (SIT) is an environmentally safe insect pest management method that has been implemented against several tephritids and other pest insects of economic significance. The efficacy of SIT relies upon sterile males outcompeting field males to copulate with field females, which then fail to reproduce, resulting in suppression of pest populations. Mass production and sterilisation by irradiation can adversely affect several male fly traits. SIT has also been developed and deployed for the control of Queensland fruit fly, Bactrocera tryoni (Froggatt), Australia’s most significant horticultural pest species. Research has shown that bacteria play an important role in tephritid biology, and some bacterial isolates can improve performance traits, including mating competitiveness. However, little is known about the application of symbiotic bacteria in enhancing tephritid performance in SIT operational programs, and this is particularly the case for B. tryoni. Symbiotic bacteria supplied to mass-reared fruit flies may help overcome some of these issues. However, the effects of tephritid ontogeny, sex, diet and irradiation on their microbiota are not well known. The aim of this PhD was to establish the diversity and abundance of bacterial symbionts in the gut of B. tryoni collected from different laboratory and field environments. Overall my research contributes to the wider research effort on the microbiota of tephritid pest fruit flies. Recent advances in sequencing technology have enabled more insights into the diversity and dynamics gut bacterial communities of insects and the roles they play in insect development. Gut bacteria have been demonstrated to improve the performance of tephritid fruit flies and thus are a promising target in improving the sterile insect technique used in tephritid fruit fly management.