Virulence of Beauveria sp. and Metarhizium sp. fungi towards fall armyworm (Spodoptera frugiperda).

The development of effective pest management strategies for Spodoptera frugiperda is a high priority for crop protection across its invasive ranges. Here, we examined six Beauveria and five Metarhizium fungal isolates against this pest. Two Beauveria isolates (B-0571, B-1311) induced high mortality toward 3rd and 6th instar caterpillars and adults. For B-0571 mortality was 82.81 ± 5.75%, 61.46 ± 6.83%, and 93.75 ± 3.61%, and 73.72 ± 2.51%, 71.88 ± 5.41%, and 97.92 ± 2.08% for B-1311, with deaths in caterpillars largely occurring under 24 h (3rd instar control 0.74 ± 0.33%, B-0571 73.96 ± 7.85% and B-1311 62.08 ± 3.67%; 6th instar control 0%, B-0571 66.67% ± 11.02% and B-1311 62.5% ± 9.55%). Infection from both Beauveria isolates fully prevented reproduction in surviving S. frugiperda females. In contrast, all five Metarhizium isolates tested and the remaining four Beauveria isolates exhibited lower virulence. The discovery of two highly virulent Beauveria fungal isolates to S. frugiperda opens avenues to develop novel biological control tools against this highly invasive pest.

Pre-release dietary supplements of methoprene and raspberry ketone increase field abundance of sterile Queensland fruit flies (Diptera: Tephritidae).

The sterile insect technique (SIT) is a sustainable pest management tool based on the release of millions of sterile insects that suppress reproduction in targeted populations. Success of SIT depends on survival, maturation, dispersal, and mating of released sterile insects. Laboratory and field cage studies have demonstrated that dietary supplements of methoprene and raspberry ketone (RK) promote sexual maturation of adult Queensland fruit fly, Bactrocera tryoni (Froggatt), and may hence shorten the delay between release and maturity in the field. We investigated the effects of methoprene and RK dietary supplements on field abundance of sexually mature sterile Q-flies relative to untreated flies fed only sugar and yeast hydrolysate before release at 2 d of age. Compared with untreated flies, more methoprene- and RK-treated flies were recaptured in cuelure traps to which only sexually mature males are attracted. At distances of 100 and 200 m from the release point, recapture rates were higher for methoprene- and RK-treated flies than for untreated flies, but at 300 m recapture rates were low and were similar for treated and untreated flies. Rainfall, relative humidity, wind speed, and wind direction did not affect recapture rates, but temperature was positively correlated with recapture rates for all treatments. There was a strong correlation between the number of sterile and wild flies caught in traps, indicating co-location in the field. Dietary supplements of methoprene and RK can substantially increase abundance of sexually mature sterile male Q-flies in the field following release as 2-d-old immature adults.

Effect of Chilling on Quality Control Parameters of Sterile Queensland Fruit Fly (Diptera: Tephritidae).

Queensland fruit fly (Q-fly), Bactrocera tryoni (Froggatt), presents a major threat to Australian fruit production and trade. The sterile insect technique (SIT) is increasingly employed to manage Q-fly. Quality of sterile males released in SIT programs, and hence program efficacy, can be affected by pre- and post-production processes, such as mass rearing, packing, irradiation, transportation, and release. Given long distances from rear-out facilities to release sites, adult flies are usually chilled to reduce metabolism and stress during transportation. To guide SIT procedures, it is important to understand the impact of such practices on performance of sterile Q-fly. The present study assesses the effect of chilling temperature and exposure period on quality parameters of sterile Q-fly. We considered the effects of two temperature regimes (4 and 6°C) and six exposure periods (0, 1, 2, 4, 6, and 12 h) on chill-coma recovery time, flight ability, survival under nutritional stress, and longevity of both males and females. Flies chilled at 4°C took longer to recover than that those chilled at 6°C. Flight ability, survival under nutritional stress, and longevity all decreased as chilling period increased but did not differ between the two tested temperatures. We recommend that periods of chilling during transportation from rear-out facilities to release sites be minimized in order to retain quality of sterile Q-fly and that increased release rates be considered when longer chilling periods are required.

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.

Electrophysiological Responses to Cuelure of Raspberry Ketone-Fed Queensland Fruit Flies.

The sterile insect technique (SIT) and male annihilation technique (MAT) are important tools for the control of Queensland fruit fly (Q-fly), Bactrocera tryoni (Froggatt) (Diptera: Tephritidae), a major insect pest of horticultural crops in Australia. In MAT, mature Q-fly males are attracted to a toxic bait using Cuelure, a synthetic analog of raspberry ketone (RK). Substantial improvements in control could be achieved by simultaneous use of SIT and MAT, but this requires suppression of the Cuelure response in released sterile flies. Recent studies report that prerelease feeding with RK during the first 48 h after emergence can reduce the response of mature Q-fly males to Cuelure, but the mechanism underpinning this is unknown. Here, to test whether reduced sensory sensitivity to Cuelure is involved, we evaluated the effects of RK supplements, adult diet (yeast-supplemented diet throughout adult stage vs yeast-supplemented diet only for 48 h), and age on electroantennogram (EAG) and electropalpogram (EPG) responses of Q-flies to Cuelure stimuli. EAG responses did not vary with RK supplements, sex, or age of Q-flies fed yeast-supplemented diet throughout the adult stage, but the responses of Q-flies fed other diet regime decreased with age. EPG responses of both sexes of Q-flies were affected by RK supplements, age, and their interaction, but without patterns that might indicate reduced maxillary palp response of RK supplemented flies to Cuelure. Our findings do not support the hypothesis that reduced Cuelure response of male Q-flies fed RK supplements is explained by reduced electrophysiological response in antennae or maxillary palps.

Artificial Larval Diet Mediates the Microbiome of Queensland Fruit Fly.

Larval diets used for artificial rearing can have a significant effect on insect biology. The Queensland fruit fly (aka "Qfly"), Bactrocera tryoni (Froggatt) (Diptera: Tephritidae), is one of the greatest challenges for fruit growers in Australia. The sterile insect technique (SIT) is being developed to manage outbreaks in regions that remain free of Qfly and to reduce populations in regions where this species is endemic. Factory scale rearing is essential for SIT; however, artificial larval diets are known to affect the microbiome of Qfly, which may then affect fly performance. In this study, high-throughput Illumina sequencing was used to assess the Qfly microbiome in colonies reared, for five generations from nature, on two common artificial diets (carrot and gel). At generation five (G5), the microbiome was assessed in larvae, pupae, adult males and adult females and standard fly quality control parameters were assessed together with additional performance measures of mating propensity and survival under nutritional stress. At the genus level, bacterial communities were significantly different between the colonies reared on the two larval diets. However, communities converged at Phyla to family taxonomic levels. Bacterial genera of Morganella, Citrobacter, Providencia, and Burkholderia were highly abundant in all developmental stages of Qfly reared on the gel diet, when compared to the carrot diet. Despite abundance of these genera, a greater percentage of egg hatching, heavier pupal weight and a higher percentage of fliers were found in the Qfly reared on the gel diet. Mating propensity and survival under nutritional stress was similar for adult Qfly that had been reared on the two larval diets. Overall, our findings demonstrate that the artificial larval diet strongly influences the microbiome and quality control measures of Qfly, with likely downstream effects on performance of flies released in SIT programs.

Effects of fatty acids and vitamin E in larval diets on development and performance of Queensland fruit fly.

Tephritid fruit flies are commonly reared on artificial larval diets for laboratory studies and for sterile insect technique pest management programs. While significant effort has been invested in developing artificial larval diets, surprisingly little is known about the specific nutritional requirements of tephritid flies. Recently developed gel larval diets have provided new opportunities for nutritional studies in Queensland fruit fly, Bactrocera tryoni ('Q-fly'). Wheat germ oil (WGO) is the main source of fatty acids and vitamin E in this diet, and is key for production of high-quality adults. To identify the importance of nutritional components of WGO for Q-fly productivity and quality, linoleic, linolenic, oleic and palmitic fatty acids as well as α-tocopherol (vitamin E) were included in the diet individually and in combination. Diets that included all of the tested fatty acids or just unsaturated fatty acids performed as well as diets containing WGO in most quality control parameters except fecundity, and addition of vitamin E reduced the pupal productivity. Considering individual fatty acids, larval diets containing only linolenic acid produced adults with higher percentage of fliers than did larval diets containing only palmitic acid or oleic acid. Compared with diets containing WGO, nutritional requirements for egg production in Q-fly were not entirely met by either grouped fatty acids or individual polyunsaturated, monounsaturated or saturated fatty acids, however, diets containing linoleic acid alone produced more eggs than any other fatty acid. The present study is a significant advance in understanding of the role of fatty acids as a component of WGO in larval diet in meeting the needs of developing Q-fly for somatic performance, but highlight also that other, untested, components of WGO appear to be important for reproduction.

Larval foraging decisions in competitive heterogeneous environments accommodate diets that support egg-to-adult development in a polyphagous fly.

In holometabolous insects, larval nutrition is a key factor underpinning development and fitness. Heterogeneity in the nutritional environment and larval competition can force larvae to forage in suboptimal diets, with potential downstream fitness effects. Little is known about how larvae respond to competitive heterogeneous environments, and whether variation in these responses affects current and next generations. Here, we designed nutritionally heterogeneous foraging arenas by modifying nutrient concentration, where groups of the polyphagous fruit fly Bactrocera tryoni could forage freely at various levels of larval competition. Larval foraging preferences were highly consistent and independent of larval competition, with greatest foraging propensity for high (100%) followed by intermediate (80% and 60%) nutrient concentration diets, and avoidance of lower concentration diets (less than 60%). We then used these larval preferences (i.e. 100%, 80% and 60% diets) in fitness assays in which larvae competition was maintained constant, and showed that nutrient concentrations selected by the larvae in the foraging trials had no effect on fitness-related traits such as egg hatching and pupation success, adult flight ability, sex ratio, percentage of emergence, nor on adult cold tolerance, fecundity and next-generation pupal weight. These results support the idea that polyphagous species can exploit diverse hosts and nutritional conditions with minimal fitness costs to thrive in new environments.

Comparison of Gel Larval Diet With Traditional Lucerne Chaff and Carrot Solid Diets for Rearing of Queensland Fruit Fly (Diptera: Tephritidae).

Sterile insect technique (SIT) for Queensland fruit fly, Bactrocera tryoni Froggatt, Australia's most economically damaging fruit fly species, is currently undergoing a major renewal and expansion. SIT relies on efficient and economical mass-rearing procedures that produce high-quality flies. Two solid larval diets, carrot and lucerne chaff, have traditionally been used to rear Queensland fruit fly. Recently, a gel larval diet has been developed to eliminate biological bulking agents from the mass-rearing process, but to date, there has been no direct comparison of gel larval diet with traditional solid diets. In the present study, the performance of flies reared on gel larval diet was compared with the performance of flies reared on carrot and lucerne chaff diets. In addition, to investigate whether the performance of reared flies depends on ancestral diet as well as tested diet, we sourced eggs from a colony maintained on carrot diet and from a colony maintained on a lucerne chaff diet. Overall, the gel diet was as good or better than the solid diets in all quality control parameters, including, egg-larval duration, pupal number, pupal recovery, adult emergence, percentage of fliers, and rate of fliers. Of note, larvae developed faster and pupated more synchronously on the gel diet than on either of the solid diets. At the loading densities used, gel and carrot diets produced less waste than lucerne chaff diet. Gel diets offer a rearing solution for Queensland fruit fly that eliminates biological bulking agents and yields faster and more synchronous larval development without compromising productivity or quality.

Canola Oil as an Economical Lipid Source in Gel Larval Diet for Queensland Fruit Fly.

A new sterile insect technique (SIT) program is currently being developed for management of the Queensland fruit fly, Bactrocera tryoni (Froggatt) (Diptera: Tephritidae), the most costly challenge to Australian horticulture in the eastern states. SIT relies on cost-effective mass production of millions of high-quality flies. A recently developed gel larval diet has proven effective, enabling production of large numbers of high-quality flies, but includes some costly ingredients. With a basic diet now available, current research focuses on refining the formulation to deliver a more economical diet. Wheat germ oil (WGO) is the main lipid source in the current Queensland fruit fly gel diet, but is a particularly expensive ingredient and has limited availability. To identify an oil that maintains high productivity and fly quality at reduced cost, the present study assessed production and performance of Queensland fruit flies reared on gel larval diets containing the WGO that was used in previous studies (WGO/M), an alternative WGO (WGO/A), sunflower oil, rice bran oil, and canola oil. Diets containing canola oil ($5.24/liter) performed as well as diets with WGO/M ($116/liter) in terms of parental egg hatch, pupal number, pupal weight, adult emergence, percentage and rate of fliers, sex ratio, fecundity, and fertility (F1 egg hatch), offering a remarkably cost-effective alternative. Costs of oil in Queensland fruit fly production are reduced by ca. 95% per 1,000 flight capable adults ('fliers'). Substantial savings may be made in Queensland fruit fly mass rearing by substituting WGO/M with canola oil in gel larval diets without compromising productivity.

Effects of Wheat Germ Oil Concentration in Gel Larval Diets on Production and Quality of Queensland Fruit Fly, Bactrocera tryoni (Diptera: Tephritidae).

Queensland fruit fly ('Q-fly'), Bactrocera tryoni (Froggatt) (Diptera: Tephritidae), is the most destructive insect pest of horticultural crops in Australia. The sterile insect technique (SIT) has attracted significant interest for sustainable management of Q-fly through the mass rearing and release of sterile flies. Cost-effective diets are required to rear Q-flies for SIT. Wheat germ oil (WGO) is the main source of fatty acids and vitamins in gel larval diets that are used to rear Q-fly but is an expensive ingredient. With the aim of reducing WGO cost in gel larval diet, we assessed performance of Q-flies reared on diets that varied in WGO content. Q-fly larvae were reared on two diets that differed mainly in yeast composition ('gel diet 2006' and 'gel diet 2009') and contained 0, 0.03, 0.07, 0.11, 0.15, or 1% WGO to identify reduced concentrations of WGO (and hence cost) without compromising productivity or quality. Diets containing WGO outperformed diets without WGO in development rate, pupal number, adult emergence, percentage of fliers, rate of fliers, and fecundity. Concentrations of 0.11% and above provided full benefit in gel diet 2006 (original formulation 0.15%), and concentrations of 0.15% (original formulation 1.0%) and above provided full benefit in gel diet 2009, and for both diets, a concentration-dependent decline in fly performance resulted from lower doses. Savings can be made in gel diets for mass rearing of Q-fly without compromising productivity by reducing WGO concentration.

Changes in nutritional composition of soybean seed caused by feeding of pentatomid (Hemiptera: Pentatomidae) and alydid bugs (Hemiptera: Alydidae).

Changes in protein, lipid, and carbohydrate content, and the weight loss of soybean seeds caused by the feeding of 6- to 7-d-old unmated male adults of the pentatomids Peizodorous hybneri (Gmelin) and Halymorpha halys (Stål), and an alydid, Riptortus pedestris (F.), were examined in the laboratory. Our goals were to determine which species had the greatest capacity to damage soybean seed and to measure the effect of that damage on the nutritional composition of soybean seed. Individuals of the three species were provided with a preweighed dry soybean seed and allowed to feed for 0, 4, 8, 12, 16, 20, and 24 d, after which the remaining seed was analyzed for any change in weight, protein, lipid, and carbohydrate concentration. Lipids, carbohydrates, and seed weights were reduced by bug feeding, and the reduction was directly proportional to feeding duration. H. halys was found to be the most voracious feeder, reducing soybean seed weight by 42% after 24 d of feeding. There was a significant interaction between species and feeding duration for changes in nutritional components. Seeds fed on by H. halys had the highest incremental increase in protein content (13%) after 24 d of feeding, followed by those fed on by R. pedestris and P. hybneri. However, carbohydrates and lipid content of the soybean seeds fed by the tested insect species were found to decrease significantly. Soybean pods at mature stages remain in the field for a long period, and findings of our study suggest that longer exposure of the mature soybean pods to these pest species in the field may lead to low-quality seeds and lower yields, and may even affect the germination potential of the seeds.

Effect of aggregation pheromone trap of Riptortus pedestris (Hemiptera: Alydidae) on the distribution and composition of its egg parasitoids.

The aggregation pheromone of Riptortus pedestris (F.) (Hemiptera: Alydidae) is known to attract its egg parasitoids Ooencyrtus nezarae Ishii (Hymenoptera: Encyrtidae) and Gryon japonicum (Ashmead) (Hymenoptera: Scelionidae). Distribution and composition of these egg parasitoids were compared in a soybean field in the presence and absence of aggregation pheromone-baited traps for two consecutive years. Three traps were installed on the edge of a soybean field, first without and then with aggregation pheromone in a before-after design, and changes in parasitism were assessed on nonviable host eggs released at distances of 9, 18, 27, and 36 m from the trap. A significant interactive effect of pheromone installation and distance was found in egg parasitism by both parasitoids. Parasitism on eggs recovered from 9 m away was up to 2.2 and 3.2 times higher in presence of pheromone than without pheromone for O. nezarae and G. japonicum, respectively. Parasitism by O. nezarae was always higher than that by G. japonicum, but their relative abundance, clutch size, and sex ratio were not affected by pheromone installation, irrespective of distance from the trap. In conclusion, aggregation pheromone increases abundance of parasitoids up to 18 m from trap installation points and does not affect the relative abundance of parasitoid species.

Interactive influence of temperature and relative humidity on egg parasitoids of Riptortus pedestris (Hemiptera: Alydidae).

Previous studies reported that of the two egg parasitoids of Riptortus pedestris (F.) (Hemiptera: Alydidae) found in Korea, Gryon japonicum (Ashmead) (Hymenoptera: Scelionidae) appears in soybean fields much earlier than Ooencyrtus nezarae Ishii (Hymenoptera: Encyrtidae). To explain this phenomenon, we evaluated the interactive influence of temperature and relative humidity (RH) on the biological attributes of these parasitoids, including adult parasitoid longevity and survival. Temperature had significant effects on all the biological attributes examined for both parasitoids, while RH only affected rates of parasitism and adult emergence. Interaction between temperature and species, but not RH and species was found to affect significantly on parasitism. G. japonicum showed higher relative increment in parasitism than O. nezarae at temperatures higher than 25 degrees C. No significant differences in progeny sex ratio were detected for either species at any temperature x humidity combination. RH had no effect on the developmental time of O. nezarae but on the developmental time of G. japonicum, which was longer at low RH. Although the biological attributes of adult parasitoids of both species showed a wide range of adaptability, but it did not explain the patterns of occurrence of these species in the field. However, G. japonicum showed greater longevity than O. nezarae at all combinations of temperature and RH and this may partially explain the seasonal pattern of occurrence of adult parasitoids in the field previously observed.