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Mass-reared sterile tephritid flies released in sterile insect technique (SIT) programmes exhibit behaviour, physiology and longevity that often differ from their wild counterparts. In the present study, video recordings of flies in laboratory cages are used to determine whether the sequential processes of mass-rearing and sterilization (using gamma radiation) that are integral to SIT affect general activity patterns of male and female Queensland fruit flies Bactrocera tryoni (Froggatt) (Diptera: Tephritidae) (‘Q-flies'). Compared with wild flies, mass-reared flies exhibit a marked reduction in overall activity, and further reduction is found after sterilization. In terms of the frequency of activities, both fertile and sterile mass-reared Q-flies fly less often and exhibit more bouts of inactivity and grooming than wild Q-flies. In addition, in terms of the duration of activities, fertile and sterile mass-reared Q-flies spend less time flying and more time walking, grooming and being inactive than wild Q-flies. Although fertile and sterile mass-reared flies are similar in other regards, sterile mass-reared flies spend more time being inactive than fertile mass-reared flies. These findings raise new questions about how changes in behaviour and activity levels may influence the performance of mass-reared sterile Q-flies in the field, as well as the physiological and metabolic processes that are involved. The frequency and duration of inactivity could provide a simple but powerful and biologically relevant test for quality in mass-rearing and SIT programs.

Queensland fruit flies, Bactrocera tryoni (Froggatt) (Diptera: Tephritidae) (‘Q-flies’) were released as sexually immature adults from a point within an orchard. Marked male Q-flies were recaptured in the trap furthest from the release point (1087 m) by 2 weeks after release, although 98.25 ± 1.04% of recaptured males were trapped

Male insects that are unable to replenish sperm supplies between matings can suffer fitness costs either because their mates are more likely to accept subsequent suitors, or because their sperm are outnumbered when females do remate. We assessed the ability of fertile and sterile (irradiated) male Queensland fruit flies, Bactrocera tryoni (‘Q-flies’), to have sperm stored by five sequential mates as well as the association between sperm depletion and female remating tendency. Sequential mates of fertile males stored similar numbers of sperm, indicating ample ability to replenish and maintain constant supplies between their once-daily mating opportunities. In contrast, sequential mates of sterile males stored progressively fewer sperm, with only trivial numbers of sperm stored by females mated by sterile males that had mated with two or more females previously. Despite the massive reduction in sperm storage by sequential mates of sterile males, fertile and sterile males were similar in their ability to induce sexual inhibition in their mates (to at least 30 days) and neither showed any decline in this ability across sequential matings. The ability of multiple-mated sterile males to induce sexual inhibition in their mates despite near or complete absence of sperm provides compelling evidence that sperm abundance plays no role in the induction of sexual inhibition in this species.

The current study is an important step toward calibrating, validating, and improving irradiation methods used for Bactrocera tryoni (Froggatt) sterile insect technique (SIT). We used routine International Atomic Energy Agency/U.S. Department of Agriculture/Food and Agriculture Organization quality control tests assessing percentage of emergence, flight ability, sex ratio, mortality under stress, reproductive sterility, and sexual competitiveness, as well as a nonstandard test of longevity under nutritional stress to assess the impact of a range of target irradiation doses (60, 65, 70, 75, and 80 Gy) on the product quality of mass reared B. tryoni used in SIT. Sterility induction remained adequate (>99.5%) for sterile male-fertile female crosses, and 100% sterility was achieved in fertile male-sterile female crosses and sterile male-sterile female crosses for each irradiation doses tested. There was significant increase in mortality under stress as irradiation dose increased, and reduced participation in mating by males irradiated at higher doses. The current target-sterilizing dose for SIT of 70-75 Gy is associated with significant reduction in fly product "quality". Our data suggest that adequate sterility and improved fly quality could be achieved through a small reduction in target sterilizing dose.

Caloric restriction (CR) has been widely accepted as a mechanism explaining increased lifespan (LS) in organisms subjected to dietary restriction (DR), but recent studies investigating the role of nutrients have challenged the role of CR in extending longevity. Fuelling this debate is the difficulty in experimentally disentangling CR and nutrient effects due to compensatory feeding (CF) behaviour. We quantified CF by measuring the volume of solution imbibed and determined how calories and nutrients influenced LS and fecundity in unmated females of the Queensland fruit fly, Bactocera tryoni (Diptera: Tephritidae). We restricted flies to one of 28 diets varying in carbohydrate:protein (C:P) ratios and concentrations. On imbalanced diets, flies overcame dietary dilutions, consuming similar caloric intakes for most dilutions. The response surface for LS revealed that increasing C:P ratio while keeping calories constant extended LS, with the maximum LS along C:P ratio of 21:1. In general, LS was reduced as caloric intake decreased. Lifetime egg production was maximized at a C:P ratio of 3:1. When given a choice of separate sucrose and yeast solutions, each at one of five concentrations (yielding 25 choice treatments), flies regulated their nutrient intake to match C:P ratio of 3:1. Our results (i) demonstrate that CF can overcome dietary dilutions; (ii) reveal difficulties with methods presenting fixed amounts of liquid diet; (iii) illustrate the need to measure intake to account for CF in DR studies and (iv) highlight nutrients rather than CR as a dominant influence on LS.

Recent laboratory studies of mass-reared flies in small cages have found that periods of just 24- or 48-h access to yeast hydrolysate can substantially enhance mating performance of mass-reared male Queensland fruit flies, Bactrocera tryoni (Froggatt) (Diptera: Tephritidae) ('Q-flies'). Using field cage tests that provide a better approximation of nature, we here investigated whether access to yeast hydrolysate for 48 h after adult emergence improves the ability of male and female mass-reared, sterile Q-flies to compete sexually with wild-type flies that had been provided continuous access to yeast hydrolysate. Mating probability of sterile males was significantly increased by 48-h access to yeast hydrolysate; sterile males provided 48-h access to yeast hydrolysate had mating probability similar to that of wild males provided continuous access to yeast hydrolysate, whereas sterile males deprived of access to yeast hydrolysate had much lower mating probability. Unlike males, access to yeast hydrolysate for 48 h did not increase mating probability of sterile female Q-flies. We instead found that wild females provided continuous access to yeast hydrolysate had higher mating probability than sterile females that did or did not have 48-h access to yeast hydrolysate. This result raises the possibility that a bisexual Q-fly strain might operate essentially as a male-only release when the flies are given access to yeast hydrolysate during a 48-h pre-release holding period. Sterile males given access to yeast hydrolysate for 48 h mated significantly earlier in the evening than wild males and, as in other recent studies, this tendency was associated with an increased tendency to mate on the trees rather than the cage walls. There was no evidence of sexual isolation in this study, as wild and sterile mass-reared flies showed no evidence of preferential mating with their own kind. Further studies are now needed to assess the potential for pre-release access to yeast hydrolysate to improve sexual performance and longevity of sterile, mass-reared, Q-flies in the field.

Feeding on yeast hydrolysate (a source of nitrogen) has a strong influence on the physiology and behaviour of the Queensland fruit fly (Q-fly), Bactrocera tryoni (Froggatt) (Diptera: Tephritidae), affecting longevity, sexual maturation, oogenesis, and mating performance. In this study, we demonstrate that access to yeast hydrolysate also influences the development of attraction to cue-lure in Q-flies. We provided virgin Q-flies various periods of access to yeast hydrolysate (continuous, 48 h, 24 h, or deprived). Attraction of males to cue-lure was increased and occurred at an earlier age when they were fed yeast hydrolysate. Males given continuous access were strongly attracted to cue-lure at a younger age (8 days after emergence), but by 12 days after emergence attraction of males given access to yeast hydrolysate for 48 h did not differ from males given continuous access. Attraction by males deprived or given just 24 h access to yeast hydrolysate was always significantly lower than those of males with continuous access. Male attraction to cue-lure was highest in the early morning. While cue-lure is most often thought of as a male attractant, virgin female Q-flies were caught in cue-lure traps at dusk at ages when they are known to be sexually mature. We suggest that cue-lure or similar natural chemicals play a role in the Q-fly mating system. γ-Irradiation used to induce sterility had no significant effect on attraction to cue-lure by Q-flies.

The mating performance field cage test is a required periodic quality-control assessment for factory-reared fruit flies used for the sterile insect technique. The FAO/IAEA/USDA guidelines for assessing fly quality state that if during tests a large proportion of flies call and mate on cage walls, away from host trees, then environmental conditions within the cage need to be adjusted and tests repeated. Here we test effects of cage design, specifically mesh colour (green, white) and addition of supplementary shade, on the mating behaviour of Queensland fruit fly (Q-fly), Bactrocera tryoni Froggatt (Diptera: Tephritidae). Observations were made over a 4-h period at dusk when these flies mate. Changes in environmental conditions in each cage over the dusk period varied with cage design. We recorded the highest proportion of matings taking place on trees as opposed to cage walls (>90%) in the unshaded white cage, the shaded white and un-shaded green cages being intermediate (ca. 70%), and the shaded green cage had the least (ca. 40%). The effects of field cage colour and supplementary shade on mating behaviour are discussed. We recommend that Q-fly field cage tests should be conducted in cages with a light coloured mesh, and that supplementary shading should only be applied if there is a need to adjust temperature and light within the cage.

Queensland fruit fly (Bactrocera tryoni; Q-fly) pupae are routinely irradiated to induce reproductive sterility in adults released in a sterile insect technique programme. Although there have been some studies of how total dose influences fly quality, dose rate has not been considered. In the present study, pupae were irradiated at a target dose range of 70‐75 Gy at dose rates of approximately 5, 7, 26, 57 and 80 Gy/min and were then subjected to routine IAEA/FAO/USDA quality control tests including emergence, flight ability, mortality under stress and sterility induction. No significant effects of dose rate were found on emergence or flight ability. Sterility induction was also found to be independent of dose rate, a result conforming to a ‘one-hit’ ionizing event hypothesis. Flies irradiated at higher dose rates suffered increased mortality under stress. This appears to stem from an increased tendency to over-shoot the target dose when irradiating at high dose rates. We recommend that, to reduce potential error in total target dose, the lowest practical dose rate be used when irradiating Q-fly pupae for use in the sterile insect technique. J. Appl. Entomol.

Dispersal of immature male and female Queensland fruit fly, Bactrocera tryoni (Froggatt) (Diptera: Tephritidae), was assessed over a period of 1 week from a single release point on three separate occasions using an array of Lynfield traps baited with cue-lure and odouriferous yellow or black sticky spheres baited with food lure (protein autolysate). Lynfield traps recaptured males; yellow or black spheres recaptured both sexes in approximately equal proportions, although at a much lower rate. As a percentage of the recapture rate for males by Lynfield traps, the mean recapture rate for yellow spheres ranged from 1.0% to 7.5% for males and 0.7% to 4.0% for females, whereas the recapture rates for black spheres ranged from 0.4% to 3.6% and 0.6% to 1.8%, respectively. The rate of recapture of sterile male flies was greater than that of unsterilised flies; this may have been due to a faster maturation rate in sterile males or because a greater proportion of them remained within the trap array rather than dispersing. There was no significant trend in recapture rate with distance from the release point to the edge of the array (88 m), except in the case of females on sticky traps where no trend was detected between 19 and 88 m. These results lend support to assumptions made about the distribution of males and females with respect to the minimum breeding density of fruit fly propagules invading a fly-free zone, and the method chosen to distribute sterile B. tryoni for the sterile insect technique.

The importance of male aggregation size for female visitation and initiation of male pheromone-calling was investigated in Bactrocera tryoni (Froggatt) (Diptera: Tephritidae) using artificial male aggregations in large laboratory cages. Female B. tryoni visited the largest aggregation more frequently than single males in association with a higher proportion of calling males, but there was no correlation between aggregation size and female visitation. Female B. tryoni had a limited capacity to perceive a difference between the number of calling males. Calling propensity of male B. tryoni was increased by the presence of conspecific males. Increased calling propensity in larger groups of male B. tryoni may be due to social facilitation of male calling behaviour. Female visitation at aggregations was only weakly associated with male calling, suggesting that aggregation size and the number of pheromone-calling males are not the only factors important in locating mates in B. tryoni, and it is possible that low-density populations could persist so long as females can encounter single males.

Background The Mediterranean fruit fly, Ceratitis capitata (Wiedemann) (Diptera: Tephritidae) is a highly invasive species now with an almost cosmopolitan distribution. Two other damaging, polyphagous and closely-related species, the marula fruit fly, Ceratitis cosyra (Walker), and the Natal fly, Ceratitis rosa Karsch, are not established outside of sub-Saharan Africa. In this study, adult water balance traits and nutritional body composition were measured in all three species at different temperatures and levels of relative humidity to determine whether tolerance of water stress may partially explain their distribution. Results Adult C. capitata exhibited higher desiccation resistance than C. rosa but not C. cosyra. Desiccation resistance of C. capitata was associated with lower rates of water loss under hot and dry conditions, higher dehydration tolerance, and higher lipid reserves that were catabolised during water stress. In comparison with C. capitata, C. cosyra and C. rosa lost water at significantly higher rates under hot, dry conditions, and did not catabolise lipids or other sources of metabolic water during water stress. Conclusions These results suggest that adult physiological traits permitting higher tolerance of water stress play a role in the success of C. capitata, particularly relative to C. rosa. The distribution of C. cosyra is likely determined by the interaction of temperature with water stress, as well as the availability of suitable hosts for larval development. Electronic supplementary material The online version of this article (doi:10.1186/s12983-016-0147-z) contains supplementary material, which is available to authorized users.

Macroinvertebrate assemblages can vary with season as a result of changing environmental conditions and phenology of individual species. The objective of this study was to determine whether results from the South African Scoring System version 5 (SASS5) are affected by season. The standard SASS5 protocol was used to sample three sites on the Skeerpoort River, South Africa, in autumn, winter and spring 2013, and summer 2014. At each site, stream morphology was characterised and physico-chemical variables were measured. Two-way analysis of variance (ANOVA) was used to compare SASS indices in relation to site and season, and a one-way ANOVA was used to test the effect of season using both current and historical data. Macroinvertebrate assemblage similarity of sites was determined using non-metric multidimensional scaling ordination, followed by analysis of similarity. Whilst macroinvertebrate assemblages differed between seasons and sites, there was no seasonal variation in the values of SASS indices, but they did differ between sites. SASS5 appears to be robust to seasonal variation in macroinvertebrate assemblage and seems suitable for use throughout the year in a perennial river draining the South African highveld region. Keywords: aquatic macroinvertebrates, bioassessment, seasonal variation, water quality African Journal of Aquatic Science 2014, 39(4): 369–376

The effects of domestication and irradiation on the mating behaviour of males of Bactrocera tryoni (Froggatt) (Diptera: Tephritidae) were investigated by caging wild, laboratory-domesticated and sterile (laboratory-domesticated, gamma-irradiated) males with wild females. Mating behaviour of mass-reared males was different from that of wild males, although behaviour of wild and sterile males was similar. Mass-reared males engaged in mounting of other males much more frequently than wild and sterile males, and began calling significantly earlier before darkness. Unnatural selection pressures imposed in mass-rearing conditions may explain these changes in mass-reared male behaviour. 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 among mating partners. Despite differences in behaviour, frequency of successful copulations and mating success were similar among wild, mass-reared and sterile males.

Background Culicoides imicola Kieffer and Culicoides bolitinos Meiswinkel (Diptera: Ceratopogonidae) are both of veterinary importance, being vectors of Schmallenberg, bluetongue and African horse sickness (AHS) viruses. Within South Africa, these Culicoides species show a marked difference in their abundances according to altitude, with C. imicola highly abundant in lower altitudes, but being replaced as the dominant species by C. bolitinos in cooler, high-altitude regions. Methods The thermal physiology of field collected adults of each species was determined to evaluate whether it could account for differences in their distribution and abundance. Critical thermal maxima (CTmax) and minima (CTmin), as well as upper and lower lethal temperatures (ULT and LLT) were assessed after acclimation temperatures of 19ˌC, 24ˌC and 29ˌC. Critical thermal limits were determined using an ecologically relevant rate of temperature change of 0.06ˌC.min−1. Results Significant differences in CTmin and CTmax were found between acclimation temperatures for C. imicola and C. bolitinos. In C. bolitinos, the LLT of individuals acclimated at 24ˌC was significantly improved (LLT50 = −6.01ˌC) compared with those acclimated at the other temperatures (LLT50 = −4ˌC). Acclimation had a weak (difference in LLT50 of only 1ˌC) but significant effect on the LLT of C. imicola. When CTmin, CTmax, LLT and ULT were superimposed on daily maximum and minimum temperature records from locations where each tested Culicoides species is dominant, it was found that temperatures frequently declined below the CTmin and LLT of C. imicola at the location where C. bolitinos was dominant. Conclusions The distribution and abundance of C. imicola is likely directly constrained by their relatively poor tolerance of lower temperatures. Results for C. bolitinos suggest that the adult phase is hardy, and it is hypothesised that the thermal biology of other life stages could determine their range. Electronic supplementary material The online version of this article (doi:10.1186/1756-3305-7-384) contains supplementary material, which is available to authorized users.

Movement of tephritid flies underpins their survival, reproduction, and ability to establish in new areas and is thus of importance when designing effective management strategies. Much of the knowledge currently available on tephritid movement throughout landscapes comes from the use of direct or indirect methods that rely on the trapping of individuals. Here, we review published experimental designs and methods from mark-release-recapture (MRR) studies, as well as other methods, that have been used to estimate movement of the four major tephritid pest genera (Bactrocera, Ceratitis, Anastrepha, and Rhagoletis). In doing so, we aim to illustrate the theoretical and practical considerations needed to study tephritid movement. MRR studies make use of traps to directly estimate the distance that tephritid species can move within a generation and to evaluate the ecological and physiological factors that influence dispersal patterns. MRR studies, however, require careful planning to ensure that the results obtained are not biased by the methods employed, including marking methods, trap properties, trap spacing, and spatial extent of the trapping array. Despite these obstacles, MRR remains a powerful tool for determining tephritid movement, with data particularly required for understudied species that affect developing countries. To ensure that future MRR studies are successful, we suggest that site selection be carefully considered and sufficient resources be allocated to achieve optimal spacing and placement of traps in line with the stated aims of each study. An alternative to MRR is to make use of indirect methods for determining movement, or more correctly, gene flow, which have become widely available with the development of molecular tools. Key to these methods is the trapping and sequencing of a suitable number of individuals to represent the genetic diversity of the sampled population and investigate population structuring using nuclear genomic markers or non-recombinant mitochondrial DNA markers. Microsatellites are currently the preferred marker for detecting recent population displacement and provide genetic information that may be used in assignment tests for the direct determination of contemporary movement. Neither MRR nor molecular methods, however, are able to monitor fine-scale movements of individual flies. Recent developments in the miniaturization of electronics offer the tantalising possibility to track individual movements of insects using harmonic radar. Computer vision and radio frequency identification tags may also permit the tracking of fine-scale movements by tephritid flies by automated resampling, although these methods come with the same problems as traditional traps used in MRR studies. Although all methods described in this chapter have limitations, a better understanding of tephritid movement far outweighs the drawbacks of the individual methods because of the need for this information to manage tephritid populations.

The link between environmental temperature, physiological processes and population fluctuations is a significant aspect of insect pest management. Here, we explore how thermal biology affects the population abundance of two globally significant pest fruit fly species, Ceratitis capitata (medfly) and C. rosa (Natal fruit fly), including irradiated individuals and those expressing a temperature sensitive lethal (tsl) mutation that are used in the sterile insect technique. Results show that upper and lower lethal temperatures are seldom encountered at the field sites, while critical minimum temperatures for activity and lower developmental thresholds are crossed more frequently. Estimates of abundance revealed that C. capitata are active year-round, but abundance declines markedly during winter. Temporal autocorrelation of average fortnightly trap captures and of development time, estimated from an integrated model to calculate available degree days, show similar seasonal lags suggesting that population increases in early spring occur after sufficient degree-days have accumulated. By contrast, population collapses coincide tightly with increasing frequency of low temperature events that fall below critical minimum temperatures for activity. Individuals of C. capitata expressing the tsl mutation show greater critical thermal maxima and greater longevity under field conditions than reference individuals. Taken together, this evidence suggests that low temperatures limit populations in the Western Cape, South Africa and likely do so elsewhere. Increasing temperature extremes and warming climates generally may extend the season over which these species are active, and could increase abundance. The sterile insect technique may prove profitable as climates change given that laboratory-reared tsl flies have an advantage under warmer conditions.

International audience; Traps are used commonly to establish the presence and population density of pest insects. Deriving estimates of population density from trap data typically requires knowledge of the properties of the trap (e.g. active area, strength of attraction) as well as some properties of the population (e.g. diffusion rate). These parameters are seldom exactly known, and also tend to vary in time, (e.g. as a result of changing weather conditions, insect physiological condition). We propose using a set of traps in such a configuration that they have different rate of trapping the insects. The properties of the traps and the characteristics of the population, including its density, are simultaneously estimated from the insects captured in these traps. The basic model is an advection-diffusion equation where the traps are represented via suitable advection term defined on the active area of the trap. The values of the unknown parameters of the model are derived by solving an optimization problem. Numerical simulations demonstrate the accuracy and the robustness of this method of parameter identification.

Copula duration and sperm storage patterns can directly or indirectly affect fitness of male and female insects. Although both sexes have an interest in the outcome, research has tended to focus on males. To investigate female influences, we compared copula duration and sperm storage of Queensland fruit fly females that were intact, or had been incapacitated through decapitation or abdomen isolation. We found that copulations were far longer when females had been incapacitated, indicating that constraints imposed on copula duration by intact females had been relaxed. Repeatability of copula duration for males was very low regardless of female treatment, and this is also consistent with strong female influence. Number of sperm in the spermathecae was not influenced by female treatment, suggesting that female abdominal ganglia control the transport of sperm to these long-term storage organs. However, more sperm were found in the ventral receptacles of incapacitated females compared to intact females. Overall, results implicate cephalic ganglia in regulation of copula duration and short-term sperm storage in the ventral receptacle and abdominal ganglia in regulation of long-term sperm storage in the spermathecae.

The molecular details of hypoxia-induced cellular responses have been difficult to identify since there is as yet no known oxygen receptor. We used cDNA microarray technology to extend our studies pertaining to these molecular details in human hepatocellular carcinoma (Hep3B) cells that produce erythropoietin (Epo) in response to hypoxia. Of approximately 1200 genes in the array, those associated with integrin-linked kinase (ILK), fibronectin precursor and glycogen synthase kinase-3beta (GSK-3beta) were markedly stimulated after exposure of Hep3B cells to low oxygen (1%) for 6 h. Epo, HIF-1, and von Hippel-Lindau cDNAs were measured in parallel as markers of low oxygen responses in Hep3B cells. ILK is a serine, threonine protein kinase that interacts with the cytoplasmic domains of integrin beta1 and beta3. This interaction localizes ILK to focal adhesion plaques. ILK is stimulated by cell-fibronectin interaction as well as insulin. It is regulated in a phosphatidylinositol 3-kinase dependent manner and can phosphorylate protein kinase B (PKB/AKT) and GSK-3beta. As a result of these and other activities ILK has been shown to affect anchorage-independent cell survival, cell cycle progression and tumorigenesis in nude mice. ILK has also been implicated in the Wnt pathway and as a critical target in PTEN-dependent tumor therapies. To our knowledge this is the first report implicating the ILK pathway in low oxygen responses. Other genes identified as a result of the microarray analysis not previously known to change as a result of low oxygen treatment were elongation factor-1alpha, glycyl-tRNA synthetase, and laminin receptor protein-1. These findings were all corroborated by RT-PCR assays and in some instances Western blot analysis.