Shaping the Microbial Landscape: Parasitoid-Driven Modifications of Bactrocera dorsalis Microbiota.

Koinobiont endoparasitoids regulate the physiology of their hosts through altering host immuno-metabolic responses, processes which function in tandem to shape the composition of the microbiota of these hosts. Here, we employed 16S rRNA and ITS amplicon sequencing to investigate whether parasitization by the parasitoid wasps, Diachasmimorpha longicaudata (Ashmaed) (Hymenoptera: Braconidae) and Psyttalia cosyrae (Wilkinson) (Hymenoptera: Braconidae), induces gut dysbiosis and differentially alter the gut microbial (bacteria and fungi) communities of an important horticultural pest, Bactrocera dorsalis (Hendel) (Diptera: Tephritidae). We further investigated the composition of bacterial communities of adult D. longicaudata and P. cosyrae to ascertain whether the adult parasitoids and parasitized host larvae share microbial taxa through transmission. We demonstrated that parasitism by D. longicaudata induced significant gut perturbations, resulting in the colonization and increased relative abundance of pathogenic gut bacteria. Some pathogenic bacteria like Stenotrophomonas and Morganella were detected in both the guts of D. longicaudata-parasitized B. dorsalis larvae and adult D. longicaudata wasps, suggesting a horizontal transfer of microbes from the parasitoid to the host. The bacterial community of P. cosyrae adult wasps was dominated by Arsenophonus nasoniae, whereas that of D. longicaudata adults was dominated by Paucibater spp. and Pseudomonas spp. Parasitization by either parasitoid wasp was associated with an overall reduction in fungal diversity and evenness. These findings indicate that unlike P. cosyrae which is avirulent to B. dorsalis, parasitization by D. longicaudata induces shifts in the gut bacteriome of B. dorsalis larvae to a pathobiont-dominated community. This mechanism possibly enhances its virulence against the pest, further supporting its candidacy as an effective biocontrol agent of this frugivorous tephritid fruit fly pest.

Effects of Physiological Status and Environmental Factors on the Lure Responses of Three Pest Fruit Fly Species (Diptera: Tephritidae).

Biotic and abiotic factors influence how insects respond to stimuli. This can make it challenging to interpret captures in traps used to monitor pest abundance in management programmes. To address this, the lure response of three pest fruit flies (Diptera: Tephritidae) was evaluated in a semi-field setting with respect to several physiological and environmental factors. Using standardised methods with known fly numbers in field cages, the response to Biolure (food-based lure) was evaluated for Ceratitis capitata, Ceratitis cosyra and Bactrocera dorsalis. Response to the male lures was tested: E.G.O PheroLure for C. capitata and C. cosyra, Trimedlure for C. capitata, and methyl eugenol for B. dorsalis. The physiological variables evaluated were fly age, sex, weight, and total body nutritional composition. The environmental effects of temperature, relative humidity and light intensity were also assessed. Protein-deprived adults responded more strongly to Biolure. The response to Biolure was not sex-specific. Fly age influenced the response of all species to all tested lures. However, this effect was species and lure specific. Temperature was the most influential environmental factor, with response generally increasing with temperature. Lower thresholds for lure response, despite the proximity of responsive flies, range from 12.21 to 22.95 °C depending on the species and lure tested. These results indicate that trapping systems and management activity thresholds must take physiological and environmental variation into account to increase their accuracy.

Addition of Selected Plant-Derived Semiochemicals to Yellow Sticky Traps Does Not Improve Citrus Psyllid Captures.

Fast and effective monitoring and surveillance techniques are crucial for the swift implementation of control methods to prevent the spread of Huanglongbing, a devastating citrus disease, and its invasive psyllid vector, Asian citrus psyllid, Diaphorina citri, into South Africa, as well as to control the native vector, African citrus triozid, Trioza erytreae. Monitoring for citrus psyllid pests can be improved by using semiochemical odorants to augment already visually attractive yellow sticky traps. However, environmental variables such as temperature and humidity could influence odorant release rates. Five field cages were used to test the ability of a selection of odorants to improve yellow sticky trap efficacy in capturing citrus psyllids. Environmental effects on odorant loss from the dispensers were also investigated. The odorants that most improved yellow sticky trap captures in field cages were then tested under open field conditions alongside lower concentrations of those same lures. Gas chromatography-mass spectrometry was used to calculate odorant release rates as well as to determine if any contamination occurred under field conditions. None of the odorants under field cage or field conditions significantly improved psyllid capture on yellow sticky traps. Temperature influenced odorant loss, and release rate from polyethylene bulbs decreased over time. Based on these results, the use of unbaited yellow sticky traps seems to be the most effective method for monitoring of Huanglongbing vectors.

Male Mediterranean fruit flies prefer warmer temperatures that improve sexual performance.

Females and males have divergent strategies of energy investment, so the thermal preference of each sex in insects may differ because energetic conversion of metabolic reserves is dependent on temperature. We determined the thermal preference of virgin, sexually mature Mediterranean fruit flies, Ceratitis capitata, and found that males preferred a significantly higher temperature (23.8 ± 0.3 °C) than that of females (22.1 ± 0.3 °C). We then tested predictions for the difference in thermal preference related to the energetic demands of reproduction over a range of temperatures. The frequency and duration of calling bouts by male C. capitata were optimal at 26 °C. Mating propensity and latency, and copula duration, were optimal over the range of 22-28 °C. When mating occurred, temperature had little effect on the incidence of sperm storage by females, but there was a notable decline in the number stored at 28 °C. Female lifespan was highest at 18 °C, but lifetime egg production was optimal at 24 °C. These results illustrate temperature-related differences in the reproductive fitness of the sexes in C. capitata, although the optima for male traits align best with their thermal preference. They also support the theoretical prediction that insect thermal preference should be lower than the optimum for fitness.

Adult diet does not compensate for impact of a poor larval diet on stress resistance in a tephritid fruit fly.

Adult holometabolous insects may derive metabolic resources from either larval or adult feeding, but little is known of whether adult diets can compensate for deficiencies in the larval diet in terms of stress resistance. We investigated how stress resistance is affected and compensated for by diet across life stages in the marula fruit fly Ceratitis cosyra (Diptera: Tephritidae). Larvae were fed diets containing either 8% torula yeast, the standard diet used to rear this species, or 1% yeast (low protein content similar to known host fruit). At emergence, adults from each larval diet were tested for initial mass, water content, body composition, and desiccation and starvation resistance or they were allocated to one of two adult diet treatments: sucrose only, or sucrose and yeast hydrolysate. The same assays were then repeated after 10 days of adult feeding. Development on a low protein larval diet led to lower body mass and improved desiccation and starvation resistance in newly emerged adults, even though adults from the high protein larval diet had the highest water content. Adult feeding decreased desiccation or starvation resistance, regardless of the diet provided. Irrespective of larval diet history, newly emerged, unfed adults had significantly higher dehydration tolerance than those that were fed. Lipid reserves played a role in starvation resistance. There was no evidence for metabolic water from stored nutrients extending desiccation resistance. Our findings show the possibility of a nutrient-poor larval environment leading to correlated improvement in adult performance, at least in the short term.

Physiological mechanisms of dehydration tolerance contribute to the invasion potential of Ceratitis capitata (Wiedemann) (Diptera: Tephritidae) relative to its less widely distributed congeners.

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.

Effect of temperature on development of the blowfly, Lucilia cuprina (Wiedemann) (Diptera: Calliphoridae).

The blowfly Lucilia cuprina is a primary colonizer of decaying vertebrate carrion, and its development provides a temperature-dependent clock that may be used to estimate the post-mortem interval of corpses and carcasses in medicolegal forensic investigations. This study uses the development of L. cuprina raised on a substrate of chicken liver at six constant temperatures from 18 to 33 °C to calibrate a thermal accumulation model of development for forensic applications. Development was optimal near 24 °C; above this temperature, survival of post-feeding life stages was increasingly compromised, while below it, development was increasingly retarded. The lower developmental threshold (~12 °C) and thermal summation constants of L. cuprina are distinct from those reported for Lucilia sericata, verifying that it is essential to identify African Lucilia specimens accurately when using them to estimate post-mortem intervals.

Pheromonal variation and mating between two mitotypes of fall armyworm (Spodoptera frugiperda) in Africa.

In the Americas, the fall armyworm (Spodoptera frugiperda) exists in two genetically distinct strains, the corn (C) and rice (R) strains. Despite their names, these strains are not associated with host plant preferences but have been shown to vary in pheromone composition and male responses. Recently, S. frugiperda was detected in Africa as an invasive species, but knowledge about variation in strain types, pheromone composition and inter-strain mating of populations of the pest in the continent has not been fully examined. Therefore, this study aimed to investigate variations, if any in the pheromone composition of female moths, male moth responses, and mating between C and R mitotypes of S. frugiperda populations in Kenya, as well as their geographic distribution. Strains (mitotypes) of S. frugiperda were identified using mitochondrial DNA (mtDNA) markers, and their pheromonal composition determined by coupled gas chromatography-mass spectrometric (GC-MS) analysis. Male moth responses to these compounds were evaluated using GC-electroantennographic detection (EAD), electroantennogram (EAG), and wind tunnel assays. Oviposition assays were used to determine whether R and C mitotype moths could mate and produce eggs. The results showed that both the R and C mitotypes were present, and there were no statistically significant differences in their distribution across all sampled locations. Five pheromone compounds including (Z)-7-dodecenyl acetate (Z7-12:OAc), (Z)-7-tetradecenyl acetate (Z7-14:OAc), (Z)-9-tetradecenyl acetate (Z9-14:OAc), (Z)-11-tetradecenyl acetate (Z11-14:OAc) and (Z)-11-hexadecenyl acetate (Z11-16:OAc), were detected in the pheromone glands of female moths of both mitotypes, with Z9-14:OAc being the most abundant. The relative percentage composition of Z9-14:OAc was similar in both mitotypes. However, the R mitotype had a 2.7 times higher relative percentage composition of Z7-12:OAc compared to the C mitotype moth, while the C mitotype moth had a 2.4 times higher relative percentage composition of Z11-16:OAc than the R mitotype moth. Male moths of both mitotypes exhibited similar responses to the pheromone compounds, showing the strongest responses to Z9-14:OAc and Z7-12:OAc in electrophysiological and behavioural assays. There was mating between R and C mitotypes with egg production comparable to mating within the same mitotype. Our results revealed that differences between the two S. frugiperda mitotypes are characterized by female moth pheromone composition rather than male moth responses to the pheromones, and that this does not prevent hybridisation between the mitotypes, which may have implications for their management.

Larval and adult diet affect phenotypic plasticity in thermal tolerance of the marula fly, Ceratitis cosyra (Walker) (Diptera: Tephritidae).

Introduction: Temperature fluctuations are important for the distribution and survival of insects. Rapid hardening, a type of phenotypic plasticity, is an adaptation that can help individuals better tolerate lethal temperatures because of earlier exposure to a sublethal but stressful temperature. Nutrition and sex are also known to influence a species ability to tolerate thermal stress. This study determined the effects of larval diet, adult diet, sex and hardening on the thermal tolerance of Ceratitis cosyra (Walker) (Diptera: Tephritidae) at lower and upper lethal temperatures. Methods: Larvae were raised on either an 8% torula yeast (high) or a 1% torula yeast (low) larval diet and then introduced to one of three dietary regimes as adults for thermal tolerance and hardening assays: no adult diet, sugar only, or sugar and hydrolysed yeast diet. Flies of known weight were then either heat- or cold-hardened for 2 hours before being exposed to a potentially lethal high or low temperature, respectively. Results: Both nutrition and hardening as well as their interaction affected C. cosyra tolerance of stressful temperatures. However, this interaction was dependent on the type of stress, with nutrient restriction and possible adult dietary compensation resulting in improved cold temperature resistance only. Discussion: The ability of the insect to both compensate for a low protein larval diet and undergo rapid cold hardening after a brief exposure to sublethal cold temperatures even when both the larva and the subsequent adult fed on low protein diets indicates that C. cosyra have a better chance of survival in environments with extreme temperature variability, particularly at low temperatures. However, there appears to be limitations to the ability of C. cosyra to cold harden and the species may be more at risk from long term chronic effects than from any exposure to acute thermal stress.

Responses of the fall armyworm (Spodoptera frugiperda) to different host plants: Implications for its management strategy.

BACKGROUND: The selection of suitable host plants for oviposition is critical for herbivorous insects to maximise survival of their offspring. Olfaction plays an important role in this process. However, little is known about how olfaction shapes the interaction between the fall armyworm (Spodoptera frugiperda) and host plants. In this study, we tested the hypothesis that olfaction guides the host selection process in the fall armyworm using oviposition and wind tunnel bioassays. RESULTS: In no-choice and dual-choice assays, female moths oviposited on all seven host plants that were tested (maize, sorghum, wheat, bean, cowpea, tomato and cabbage). However, in multiple-choice assays, no eggs were deposited on cowpea and cabbage. We found that maize, sorghum and wheat were most preferred for oviposition, whereas cowpea was least preferred. Wind tunnel assays confirmed these divergent oviposition preferences, with maize, sorghum and wheat odours being the most attractive. Gas chromatography-mass spectrometry analysis followed by random forest classification identified terpenes as the potential host-plant attractants. CONCLUSION: Our results improve our understanding of the chemical ecology of the fall armyworm and suggest that some of these host plants could offer potential for use in an intercropping strategy to manage S. frugiperda. © 2022 Society of Chemical Industry.

A protocol for evaluating the entomological impact of larval source reduction on mosquito vectors at hotel compounds in Zanzibar.

There is an increasing awareness of the association between tourism activity and risks of emerging mosquito-borne diseases (MBDs) worldwide. In previous studies we showed that hotels in Zanzibar may play an important role in maintaining residual foci of mosquito vectors populations of public health concern. These findings indicated larval sources removal (LSR) interventions may have a significant negative impact on vector communities. However, a thorough analysis of the response vector species to potential LSM strategies must be evaluated prior to implementation of a large-scale area-wide control campaign. Here we propose a protocol for evaluation of the impact of LSR against mosquito vectors at hotel settings in Zanzibar. This protocol is set to determine the efficacy of LSR in a randomized control partial cross-over experimental design with four hotel compounds representing the unit of randomization for allocation of interventions. However, the protocol can be applied to evaluate the impact of LRS in more than four sites. Proposed interventions are active removal of disposed containers, and installation of water dispenser to replace single use discarded plastic water bottles, which were identified as the most important source of mosquitoes studied hotels. The ideal time for allocating intervention to the intervention arms the dry season, when the mosquito abundance is predictably lower. The possible impact of interventions on mosquito occurrence and abundance risks is then evaluated throughout subsequent rainy and dry seasons. If an appreciable reduction in mosquito abundance and occurrence risks is observed during the trial period, intervention could be extended to the control arm to determine whether any potential reduction of mosquito density is reproducible. A rigorous evaluation of the proposed LRS interventions will inspire large scale trials and provide support for evidence-based mosquito management at hotel facilities in Zanzibar and similar settings.

Selection on female reproductive schedules in the marula fly, Ceratitis cosyra (Diptera: Tephritidae) affects dietary optima for female reproductive traits but not lifespan.

Introduction: A changing environment can select on life-history traits and trade-offs in a myriad of ways. For example, global warming may shift phenology and thus the availability of host-plants. This may alter selection on survival and fertility schedules in herbivorous insects. If selection on life-histories changes, this may in turn select for altered nutrient intake, because the blend of nutrients organisms consume helps determine the expression of life-history traits. However, we lack empirical work testing whether shifts in the timing of oviposition alter nutrient intake and life-history strategies. Methods: We tested in the marula fruit fly, Ceratitis cosyra, how upward-selection on the age of female oviposition, in comparison with laboratory adapted control flies, affects the sex-specific relationship between protein and carbohydrate intake and life-history traits including lifespan, female lifetime egg production and daily egg production. We then determined the macronutrient ratio consumed when flies from each selection line and sex were allowed to self-regulate their intake. Results: Lifespan, lifetime egg production and daily egg production were optimised at similar protein to carbohydrate (P:C) ratios in flies from both selection lines. Likewise, females and males of both lines actively defended similar nutrient intake ratios (control =1:3.6 P:C; upward-selected = 1:3.2 P:C). Discussion: Our results are comparable to those in non-selected C. cosyra, where the optima for each trait and the self-selected protein to carbohydrate ratio observed were nearly identical. The nutrient blend that needs to be ingested for optimal expression of a given trait appeared to be well conserved across laboratory adapted and experimentally selected populations. These results suggest that in C. cosyra, nutritional requirements do not respond to a temporal change in oviposition substrate availability.

Friend or Foe: Symbiotic Bacteria in Bactrocera dorsalis-Parasitoid Associations.

Parasitoids are promising biocontrol agents of the devastating fruit fly, Bactrocera dorsalis. However, parasitoid performance is a function of several factors, including host-associated symbiotic bacteria. Providencia alcalifaciens, Citrobacter freundii, and Lactococcus lactis are among the symbiotic bacteria commonly associated with B. dorsalis, and they influence the eco-physiological functioning of this pest. However, whether these bacteria influence the interaction between this pest and its parasitoids is unknown. This study sought to elucidate the nature of the interaction of the parasitoids, Fopius arisanus, Diachasmimorpha longicaudata, and Psyttlia cosyrae with B. dorsalis as mediated by symbiotic bacteria. Three types of fly lines were used: axenic, symbiotic, and bacteria-mono-associated (Lactococcus lactis, Providencia alcalifaciens, and Citrobacter freundii). The suitable stages of each fly line were exposed to the respective parasitoid species and reared until the emergence of adult flies/parasitoids. Thereafter, data on the emergence and parasitoid fitness traits were recorded. No wasps emerged from the fly lines exposed to P. cosyrae. The highest emergence of F. arisanus and D. longicaudata was recorded in the L. lactis fly lines. The parasitoid progeny from the L. lactis and P. alcalifaciens fly lines had the longest developmental time and the largest body size. Conversely, parasitoid fecundity was significantly lower in the L. lactis lines, whereas the P. alcalifaciens lines significantly improved fecundity. These results elucidate some effects of bacterial symbionts on host-parasitoid interactions and their potential in enhancing parasitoid-oriented management strategies against B. dorsalis.

Tethered-flight performance of thermally-acclimated pest fruit flies (Diptera: Tephritidae) suggests that heat waves may promote the spread of Bactrocera species.

BACKGROUND: Thermal history may induce phenotypic plasticity in traits that affect performance and fitness. One type of plastic response triggered by thermal history is acclimation. Because flight is linked to movement in the landscape, trapping and detection rates, and underpins the success of pest management tactics, it is particularly important to understand how thermal history may affect pest insect flight performance. We investigated the tethered-flight performance of Ceratitis capitata, Bactrocera dorsalis and Bactrocera zonata (Diptera: Tephritidae), acclimated for 48 h at 20, 25 or 30 °C and tested at 25 °C. We recorded the total distance, average speed, number of flight events and time spent flying during 2-h tests. We also characterized morphometric traits (body mass, wing shape and wing loading) that can affect flight performance. RESULTS: The main factor affecting most flight traits was body mass. The heaviest species, B. dorsalis, flew further, was faster and stopped less often in comparison with the two other species. Bactrocera species exhibited faster and longer flight when compared with C. capitata, which may be associated with the shape of their wings. Moreover, thermal acclimation had sex- and species-specific effects on flight performance. Flies acclimated at 20 °C stopped more often, spent less time flying and, ultimately, covered shorter distances. CONCLUSION: Flight performance of B. dorsalis is greater than that of B. zonata and C. capitata. The effects of thermal acclimation are species-specific. Warmer acclimation temperatures may allow pest fruit flies to disperse further and faster. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Patterns of Remating Behaviour in Ceratitis (Diptera: Tephritidae) Species of Varying Lifespan.

Trade-offs between life-history traits offset the energetic costs of maintaining fitness in complex environments. Ceratitis species have been recorded to have long lifespans, which may have evolved in response to seasonal resource fluctuation. It is thus likely that reproductive patterns have evolved concomitantly as part of the trade-off between lifespan and reproduction. In this study, we investigated how reproductive patterns differ between Ceratitis cosyra (Walker) and Ceratitis capitata (Wiedemann; Diptera: Tephritidae), two species with different average and maximum lifespans. Females of both species were mated and patterns of female survival, fecundity, remating and sperm storage were tested. Ceratitis cosyra had a higher rate of survival and a lower fecundity when compared with the shorter-lived C. capitata, suggesting that both species exhibit a trade-off between lifespan and reproduction. Both species showed a similar and consistent willingness to remate, despite declines in sperm storage, suggesting that sperm alone does not fully inhibit remating. As expected, C. cosyra transferred high numbers of sperm during the first mating. However, sperm stores declined unexpectedly by 14 days. This indicates that males might transfer large ejaculates as a nuptial gift, that females then later degrade as a source of nutrients. Large declines in sperm storage may also indicate that females discard excess sperm stores due to the toxicity involved with storing sperm. These results do not suggest that patterns of sperm storage and remating align with lifespan and resource seasonality in these species, but a wider range of species needs to be assessed to better understand variation in Ceratitis mating systems.

Oxidative Damage Is Influenced by Diet But Unaffected by Selection for Early Age of Oviposition in the Marula Fly, Ceratitis cosyra (Diptera: Tephritidae).

The expression of life-history traits, such as lifespan or reproductive effort, is tightly correlated with the amount and blend of macronutrients that individuals consume. In a range of herbivorous insects, consuming high protein to carbohydrate ratios (P:C) decreases lifespan but increases female fecundity. In other words, females face a resource-based trade-off between lifespan and fecundity. Redox metabolism may help mediate this trade-off, if oxidative damage is elevated by reproductive investment and if this damage, in turn, reduces lifespan. Here, we test how diets varying in P:C ratio affect oxidative damage and antioxidant protection in female and male of the marula fly, Ceratitis cosyra (Diptera: Tephritidae). We use replicated lines that have been subjected to experimental evolution and differ in their lifespan and reproductive scheduling. We predicted that high fecundity would be associated with high oxidative damage and reduced antioxidant defences, while longer lived flies would show reduced damage and elevated antioxidant defences. However, higher levels of oxidative damage were observed in long-lived control lines than selection lines, but only when fed the diet promoting lifespan. Flies fed diets promoting female fecundity (1:4 and 1:2 P:C) suffered greater oxidative damage to lipids than flies fed the best diet (0:1 P:C) for lifespan. Total antioxidant capacity was not affected by the selection regime or nutrition. Our results reiterate the importance of nutrition in affecting life-history traits, but suggest that in C. cosyra, reactive oxygen species play a minimal role in mediating dietary trade-offs between lifespan and reproduction.

Differential immune responses in new and old fruit fly-parasitoid associations: Implications for their management.

The oriental fruit fly, Bactrocera dorsalis (Hendel), and marula fruit fly, Ceratitis cosyra (Walker), are major fruit-infesting tephritids across sub-Saharan Africa. Biological control of these pests using parasitic wasps has been widely adopted but with varying levels of success. Most studies investigating host-parasitoid models have focused on functional and evolutionary aspects leaving a knowledge gap about the physiological mechanisms underpinning the efficacy of parasitoids as biocontrol agents of tephritids. To better understand these physiological mechanisms, we investigated changes in the cellular immune responses of C. cosyra and B. dorsalis when exposed to the parasitic wasps, Diachasmimorpha longicaudata (Ashmaed) and Psyttalia cosyrae (Wilkinson). We found that B. dorsalis was more resistant to parasitisation, had a higher hemocyte count, and encapsulated more parasitoid eggs compared to C. cosyra, achieving up to 100% encapsulation when exposed to P. cosyrae. Exposing B. dorsalis to either parasitoid species induced the formation of a rare cell type, the giant multinucleated hemocyte, which was not observed in C. cosyra. Furthermore, compared to P. cosyrae-parasitized larvae, those of both host species parasitized by D. longicaudata had lower encapsulation rates, hemocyte counts and spreading abilities and yielded a higher number of parasitoid progeny with the highest parasitoid emergence (72.13%) recorded in C. cosyra. These results demonstrate that cellular immune responses are central to host-parasitoid interaction in tephritid fruit flies and further suggest that D. longicaudata presents greater potential as a biocontrol agent of B. dorsalis and C. cosyra in horticultural cropping systems.

High DDT resistance without apparent association to kdr and Glutathione-S-transferase (GST) gene mutations in Aedes aegypti population at hotel compounds in Zanzibar.

Global efforts to control Aedes mosquito-transmitted pathogens still rely heavily on insecticides. However, available information on vector resistance is mainly restricted to mosquito populations located in residential and public areas, whereas commercial settings, such as hotels are overlooked. This may obscure the real magnitude of the insecticide resistance problem and lead to ineffective vector control and resistance management. We investigated the profile of insecticide susceptibility of Aedes aegypti mosquitoes occurring at selected hotel compounds on Zanzibar Island. At least 100 adults Ae. aegypti females from larvae collected at four hotel compounds were exposed to papers impregnated with discriminant concentrations of DDT (4%), permethrin (0.75%), 0.05 deltamethrin (0.05%), propoxur (0.1%) and bendiocarb (0.1%) to determine their susceptibility profile. Allele-specific qPCR and sequencing analysis were applied to determine the possible association between observed resistance and presence of single nucleotide polymorphisms (SNPs) in the voltage-gated sodium channel gene (VGSC) linked to DDT/pyrethroid cross-resistance. Additionally, we explored the possible involvement of Glutathione-S-Transferase gene (GSTe2) mutations for the observed resistance profile. In vivo resistance bioassay indicated that Ae. aegypti at studied sites were highly resistant to DDT, mortality rate ranged from 26.3% to 55.3% and, moderately resistant to deltamethrin with a mortality rate between 79% to and 100%. However, genotyping of kdr mutations affecting the voltage-gated sodium channel only showed a low frequency of the V1016G mutation (n = 5; 0.97%). Moreover, for GSTe2, seven non-synonymous SNPs were detected (L111S, C115F, P117S, E132A, I150V, E178A and A198E) across two distinct haplotypes, but none of these were significantly associated with the observed resistance to DDT. Our findings suggest that cross-resistance to DDT/deltamethrin at hotel compounds in Zanzibar is not primarily mediated by mutations in VGSC. Moreover, the role of identified GSTe2 mutations in the resistance against DDT remains inconclusive. We encourage further studies to investigate the role of other potential insecticide resistance markers.

Spatio-temporal distribution of sexual calling behaviour in domesticated, sterile and wild Queensland fruit fly males under field cage conditions.

BACKGROUND: The sterile insect technique (SIT) is used in Australia to contain and eradicate outbreaks of Bactrocera tryoni (Froggatt) (Diptera: Tephritidae) in fruit-fly-free regions, and to manage populations in some endemic regions. To assess the likely impact of SIT on wild B. tryoni populations it is important to assess the colocation and synchrony of male calling between sterile and wild flies. We observed the location and timing of calling behaviours of marked mass-reared (fertile and sterile) and wild B. tryoni males in walk-in field cages. RESULTS: We found that wild males called further from the canopy centre than mass-reared (fertile or sterile) males. Mass-reared (fertile or sterile) males called earlier in the evening than wild males and, consequently, mass-reared males called when temperature and light intensity were higher than when wild males called. CONCLUSION: Male calling is a prerequisite to mating among dacine fruit flies. Therefore, our observations of spatio-temporal divergence in male calling behaviour may lead to assortative mating between mass-reared and wild B. tryoni in SIT applications. The importance of these spatio-temporal differences warrants further inquiry, with particular focus on how environmental conditions modify calling behaviour and avenues to ameliorate differences between sterile and wild flies. © 2021 Society of Chemical Industry.

Canopy distribution and microclimate preferences of sterile and wild Queensland fruit flies.

Insects tend to live within well-defined habitats, and at smaller scales can have distinct microhabitat preferences. These preferences are important, but often overlooked, in applications of the sterile insect technique. Different microhabitat preferences of sterile and wild insects may reflect differences in environmental tolerance and may lead to spatial separation in the field, both of which may reduce the control program efficiency. In this study, we compared the diurnal microhabitat distributions of mass-reared (fertile and sterile) and wild Queensland fruit flies, Bactrocera tryoni (Froggatt) (Diptera: Tephritidae). Flies were individually tagged and released into field cages containing citrus trees. We recorded their locations in the canopies (height from ground, distance from canopy center), behavior (resting, grooming, walking, feeding), and the abiotic conditions on occupied leaves (temperature, humidity, light intensity) throughout the day. Flies from all groups moved lower in the canopy when temperature and light intensity were high, and humidity was low; lower canopy regions provided shelter from these conditions. Fertile and sterile mass-reared flies of both sexes were generally lower in the canopies than wild flies. Flies generally fed from the top sides of leaves that were lower in the canopy, suggesting food sources in these locations. Our observations suggest that mass-reared and wild B. tryoni occupy different locations in tree canopies, which could indicate different tolerances to environmental extremes and may result in spatial separation of sterile and wild flies when assessed at a landscape scale.