Behavioral analysis of learning and memory in Anastrepha fraterculus

We evaluate the influence of prior exposure to artificial substrate for oviposition on learning and memory in the fruit fly Anastrepha fraterculus (Wiedemann) (Diptera: Tephritidae). Some females were previously exposed to artificial fruits made of water, agar, and blackberry [Rubus spec. (Rosaceae)] or guava [Psidium guajava L. (Myrtaceae)] pulp for 48 and 72 h. We also studied adult flies exposed for 72 h to essential oil of lemongrass [Cymbopogon citratus (DC) Stapf, Poaceae] and adult flies from larvae exposed to the oil. Control females were naive with respect to these experimental substrates. Prior experience with blackberry‐based artificial fruits resulted in an increase in the number of punctures and deposited eggs by A. fraterculus, and memory lasted for up to 72 h. On the other hand, fly behavior was independent of exposure to guava‐based substrate. Prior exposure of 1‐ or 15‐day‐old females to artificial substrate with lemongrass oil modified innate substrate selection behavior. The scent of lemongrass oil during the larval stage modified innate oviposition responses of adult A. fraterculus. The study shows that A. fraterculus females are able to learn and retain information through chemical stimuli released by both host (blackberry and guava) and non‐host (lemongrass) species, and they can use olfactory memory obtained during the larval stage to select oviposition sites.     

Combination of edible vegetable oil and artificial fruit to reduce Bactrocera dorsalis oviposition in chilli fruits

Bactrocera dorsalis (Hendel) is a major pest of a wide variety of fruits and vegetables. This study aimed to evaluate the potency of combining edible vegetable oil application and artificial fruit provision to reduce the oviposition of B. dorsalis in chilli fruits. Experiments were conducted in laboratory using a 20‐L plastic container provided with chilli fruits and in semi field using a fruit‐bearing chilli plant caged with insect screen. The laboratory test revealed that the combination of coconut oil application and artificial fruit provision significantly reduced fruit fly visits, eggs laid and the number of infested chilli fruits no matter when the chilli fruits were without prepuncture or with prepuncture. Further study using a caged chilli plant also found that this combination had significant effects on fruit fly visits and infestation in chilli fruits. These results provide an indication that coconut oil and artificial fruit can work together to reduce fruit fly attacks on chilli fruits. Combining the use of coconut oil and artificial fruit is likely to create an integrated behavioural manipulation (push and pull) of female B. dorsalis that lead to a much lower fruit fly infestation in chilli fruits.     

γ‐Octalactone,an effective oviposition stimulant of Bactrocera tryoni

Insects commonly rely on olfactory, gustatory and visual cues when deciding where to lay eggs. The olfactory cues that stimulate oviposition in the Queensland fruit fly, Bactrocera tryoni (Froggatt) (Diptera: Tephritidae), are not well understood. Here, we show that two known oviposition stimulants of the Oriental fruit fly, Bactrocera dorsalis (Hendel) (Diptera: Tephritidae)—γ‐octalactone and benzothiazole—strongly elicit aggregation and oviposition in B. tryoni. Two other known oviposition stimulants of B. dorsalis—ethyl tiglate and 1‐octen‐3‐ol—elicit aggregation but not oviposition. Highlighting species overlap, but also differences, in oviposition stimulants, these findings have practical application for mass‐rearing in which vast numbers of flies are reared for sterile insect technique programs and may also have practical application in the development of pest management and monitoring tools.     

Behavioural responses of female Queensland fruit fly, Bactrocera tryoni, to mineral oil deposits

Behavioural responses of Queensland fruit fly, Bactrocera tryoni (Froggatt) (Diptera: Tephritidae), females to fruit dipped in water and fruit dipped in 0.5% (vol/vol) aqueous emulsions of a mineral oil were determined and analysed. The mineral oil was an nC20–22 distillation fraction of the base oil used to produce an nC23 horticultural mineral oil. Females caged with oil‐treated fruit had significantly longer prelanding intervals than females caged with water‐dipped fruit. The latter was attacked immediately or shortly after being caged with flies whereas some oil‐dipped fruit was not attacked within 180 min. The percentage of landings that led to oviposition on water‐ and oil‐treated fruit were 58 and 13%, respectively, and the percentages ovipositing after probing were 74 and 25%, respectively. Likewise, average times spent probing were 7 vs. 31 s whereas average times spent ovipositing were 321 vs. 223 s. Females spent less than half as much time on oil‐treated fruit than on water‐treated fruit. Transition probabilities of rejection, when applied to the behaviour sequence indicated that oil‐treated fruits are about nine times less likely to be infested with B. tryoni.     

Oviposition responses of Queensland fruit fly (Bactrocera tryoni) to mineral oil deposits on tomato fruit

Behavioural responses of wild and laboratory‐culture females of Queensland fruit fly, Bactrocera tryoni (Froggatt) (Diptera: Tephritidae), to mineral oil deposits on tomato fruit dipped in aqueous oil emulsions were assessed in a no‐choice test and three choice tests. The oils were two commercial products used to manage plant pests and diseases, Ampol D‐C‐Tron NR and SK EnSpray 99, one distillation fraction of the base oil of the former, and four distillation fractions of the base oil of the latter. The initial and final boiling points of the fractions were equivalent to those of n‐alkanes with chain lengths of C20–22 (Ampol), and C21–C23, C22–23.5, C22–24, and C22–24.5 (SK). For both fly types in the no‐choice test, numbers of punctures and eggs per fruit declined strongly as concentrations of the nC20–22 Ampol fraction in emulsions rose from 0.25 to 2% (vol/vol). Fly type affected the extent of responses but there was no significant interaction for fly type*oil concentration. Responses of laboratory‐culture females in the choice tests also declined as concentrations of SK and the four fractions of its base oil in emulsions rose from 0 to 0.25%. The SK nC22–24 and nC22–24.5 fractions had least impact. Responses of laboratory‐culture flies to 0.5% emulsions of the nC20–22 Ampol fraction and the nC21–23 SK fraction in choice tests were not significantly different. Likewise, responses of laboratory‐culture flies to 0.5% emulsions of the two commercial products were not significantly different. Emulsifier type did not affect numbers of punctures or eggs per fruit in choice responses of laboratory‐culture flies to 0.5% emulsions of the Ampol nC20–22 fraction or 0.5% emulsions of the SK nC21–23 fraction. If the equivalence of no‐choice and choice responses in the laboratory were to hold in the field, then unsprayed ‘sacrificial’ plants would not be necessary and oil emulsions could be used as cover sprays.     

Development of Asian parasitoids in larvae Of Drosophila Suzukii feeding on blueberry and artificial diet

In just a few years, the Asian fly Drosophila suzukii has invaded several continents and has become a very serious pest of many fruit crops worldwide. Current control methods rely on chemical insecticides or expensive and labour‐intensive cultural practices. Classical biological control through the introduction of Asian parasitoids that have co‐evolved with the pest may provide a sustainable solution on condition that they are sufficiently specific to avoid non‐target effects on local biodiversity. Here, we present the first study on the development of three larval parasitoids from China and Japan, the Braconidae Asobara japonica and the Figitidae Leptopilina japonica and Ganaspis sp., on D. suzukii. The Asian parasitoids were compared with Leptopilina heterotoma, a common parasitoid of several Drosophilidae worldwide. The three Asian species were successfully reared on D. suzukii larvae in both, blueberry and artificial diet, in contrast to L. heterotoma whose eggs and larvae were encapsulated by the host larvae. All parasitoids were able to oviposit one day after emergence. Asobara japonica laid as many eggs in larvae feeding in blueberry as in artificial diet, whereas L. heterotoma oviposited more in larvae on the artificial diet and the Asian Figitidae oviposited more in larvae feeding on blueberry. Ganaspis sp. laid very few eggs in larvae in the artificial diet, suggesting that it may be specialized in Drosophila species living in fresh fruits. These data will be used for the development of a host range testing to assess the suitability of Asian parasitoids as biological control agents in invaded regions.     

An improved culturing method for opiine fruit fly parasitoids and its application to parasitoid monitoring in the field

Good culturing methods play an important role in the study of insect behavior and its application to pest management. Here, we describe and validate a new method for rearing the parasitoid wasp, Diachasmimorpha kraussii, which attacks some of the world's worst fruit fly pests and is an internationally used biological control agent. Our method differs from standard culturing approaches by presenting adult wasps with host‐infested artificial media within a “culturing bag,” which mimics a natural (fruit) oviposition substrate. In laboratory trials using wild collected D. kraussii, the culturing bag method was compared to the use of host‐infested nectarines, and a commonly used laboratory method of presenting host‐infested artificial media within Petri dishes. The culturing bag method proved to be a significant improvement on both methods, combining the advantages of high host survival in artificial media with parasitism levels that were the equivalent to those recorded using host‐infested fruits. In our field study, culturing bags infested with the Queensland fruit fly, Bactrocera tryoni, and hung in a mixed peach and nectarine orchard proved to be effective “artificial fruits” attracting wild D. kraussii for oviposition. Significantly more adult wasps were reared from the culturing bags compared to field collected fruits. This was shown to be due to higher fruit fly larval density in the bags, as similar percentage parasitism rates were found between the culturing bags and ripe fruits. We discuss how this cheap, time‐efficient method could be applied to collecting and monitoring wild D. kraussii populations in orchards, and assist in maintaining genetic variability in parasitoid laboratory cultures.     

A real‐time PCR toolbox for accurate identification of invasive fruit fly species

Significant plant pests such as fruit flies that travel with fresh produce between countries as eggs or larvae pose a great economic threat to the agriculture and fruit industry worldwide. Time‐limited and expensive quarantine decisions require accurate identification of such pests. Immature stages are often impossible to identify, making them a serious concern for biosecurity agencies. Use of COI barcoding PCR, often the only molecular identification resource, is time‐consuming. We assess the suitability of the COI barcoding region for real‐time PCR assays to identify four pest fruit fly species (Family: Tephritidae), in a diagnostic framework. These species, namely Mediterranean fruit fly (Ceratitis capitata), Queensland fruit fly (Bactrocera tryoni), African invader fly (Bactrocera invadens) and Island fly (Dirioxa pornia) each provide a different set of genetic species delimitation problems. We discuss the benefits and limitations of using a single‐gene TaqMan^? real‐time approach for such species. Our results indicate that COI‐based TaqMan^? real‐time PCR assays, in particular for genetically distinct species, provide an accurate, sensitive and rapid diagnostic tool.     

Colonization of Lutzomyia shannoni (Diptera: Psychodidae) utilizing an artificial blood feeding technique

Laboratory colonization of hematophagous insects must include an efficient method of blood feeding, preferably by artificial means. Strict rules for obtaining animal use permits, extensive animal maintenance costs, and indirect anesthesia effects on animal health warrant the development of an artificial membrane feeding technique for sand fly colonization in laboratories. An attempt was made to colonize Lutzomyia shannoni using an artificial blood feeding membrane to replace the use of live animals commonly used for sand fly blood‐feeding purposes. Lutzomyia shannoni readily fed through a pig intestine membrane exposed at an angle of 45°. However, it did not feed through a chicken skin membrane. Olfactory attractants were unable to improve blood‐feeding efficiency. Plaster of Paris was the most suitable oviposition substrate. Female L. shannoni adults laid no eggs on moist sand substrate. Sand fly adults held in groups of ten or more laid higher numbers of eggs than did individually maintained sand flies. Inclusion of the L. longipalpis oviposition hormone dodecanoic acid or the presence of previously laid eggs did not stimulate L. shannoni oviposition. The average L. shannoni egg, larval, and pupal duration were 9.3, 36.7, and 17.8 days, respectively. The addition of a 20% sugar solution improved adult female longevity. Females survived longer (14.8 days) than males (11.9 days). Lutzomyia shannoni was successfully colonized in the laboratory for up to four generations using this artificial membrane technique.     

Oviposition preference of Anthocoris nemorum and A. nemoralis for apple and pear

Anthocoris nemorum L. and Anthocoris nemoralis Fabricius (Heteroptera: Anthocoridae) are important predators of insect pests in pome fruit. Females insert their eggs in leaf tissue. The females’ choice of oviposition site is important for the subsequent distribution of nymphs on host plants. Oviposition preference for apple and pear leaves was tested in the laboratory in four experiments (experiments 1–4). In three experiments it was tested whether simulated insect damage to leaves (experiments 5 and 6) or the presence of prey (experiment 7) influenced oviposition preference. The effect of the presence of prey was only tested for A. nemorum on apple leaves. There was a highly significant anthocorid species × plant interaction for the number of eggs laid on apple and pear leaves. Anthocoris nemorum laid more eggs on apple than on pear leaves, while A. nemoralis preferred pear. Anthocoris nemorum's preference for apple increased over the 6‐week period in which experiments 1–4 were performed, from 66% to 91% eggs laid on apple leaves. No change over time in preference was found for A. nemoralis. Across experiments 1–4, the majority of A. nemorum eggs were laid near leaf margins, whereas eggs of A. nemoralis were more commonly found in the leaf centre, 5 mm or more from the margin, with a highly significant leaf region × species interaction. There was no significant difference in preference for leaf side between A. nemorum and A. nemoralis, but there was a highly significant plant × leaf side × experiment interaction. Thus, more eggs were laid on the ventral than on the dorsal side of pear leaves in experiment 4, while significantly more eggs were laid on the dorsal side of apple leaves in experiments 3 and 4. Choice tests between damaged and healthy leaves showed that A. nemorum laid significantly more eggs on the damaged leaves, while A. nemoralis preferred healthy leaves. Anthocoris nemorum showed a near‐significant preference for ovipositing on leaves with eggs of Operophtera brumata (Lepidoptera: Geometridae). The oviposition preferences found correspond to the natural distribution of these predators in apple and pear orchards. The preference of A. nemorum for leaf margins, and of A. nemoralis for the leaf centre as an oviposition site, supports earlier observations. A preference for leaf side for oviposition site has not been reported earlier. Preference for damaged leaves could help A. nemorum to locate prey in a field situation.     

Susceptibility of selected apple cultivars to the Mediterranean fruit fly

Ceratitis capitata (Wiedemann), the Mediterranean fruit fly, is one of the key pest species affecting deciduous fruit orchards along the Mediterranean coasts. Because of global warming, C. capitata is gradually spreading north and is becoming a major pest of apples. Determining the susceptibility of the main apple varieties grown in the region will serve as a cornerstone to the management of this pest. In this study, we show the results of a field and laboratory no‐choice test conducted to determine the Medfly preferences on different apple cultivars. The seven main varieties of apples (Gala, Red Delicious, Golden Delicious, Granny Smith, Kanzi, Morgen Dallago and Fuji) were tested. The results demonstrate that C. capitata lays eggs on all apple cultivars in both field and laboratory conditions. The Granny Smith, Red Delicious and Morgen Dallago varieties showed the lowest susceptibility in laboratory conditions, (0.75, 1.55, 2 oviposition punctures/fruit, respectively), with significant differences in oviposition compared to the Golden Delicious, Kanzi and Fuji (3.27, 3.31, 3.1 oviposition punctures/fruit, respectively) varieties, which were shown to be the most susceptible to Medfly attack in laboratory conditions. On the other hand, only slight and not statistically significant differences emerged from the field trials. In relation to the physico‐chemical characteristics, the apple cultivars showing the lowest susceptibility (Granny Smith, Red Delicious and Morgen Dallago) had harder peels and pulps and lower sugar contents than the most susceptible cultivars (Golden Delicious, Fuji and Kanzi). These results were also confirmed through evaluation of larval development on different varieties. In fact, Granny Smith, Red Delicious and Morgen Dallago were the three varieties that did not allow adequate larval and adult development and reduced the possibility of the emergence of a new generation.     

Oviposition preference in the oligophagous tomato fruit fly, Neoceratitis cyanescens

We assessed the role of visual and olfactory cues on oviposition preference in the oligophagous tomato fruit fly, Neoceratitis cyanescens (Bezzi) (Diptera: Tephritidae). In a field survey, we evaluated the stage of susceptibility of field‐grown tomatoes by monitoring N. cyanescens infestations from fruit‐setting up to harvest, in relation to post‐flowering time, size, and visual properties of fruit. In two‐choice laboratory experiments, we tested the degree to which females use visual and olfactory cues to select their host plant for oviposition. In addition, we investigated the ability of flies to avoid fruit already infested by conspecific eggs or larvae, and the influence of natal host fruit on oviposition preference. Neoceratitis cyanescens females preferentially lay their eggs in small yellow‐green unripe fruit (2–3.5 cm diameter, 10–21 days post‐flowering). Damage to fruit was significantly affected by brightness and size properties. In laboratory experiments, females chose to lay their eggs in bright orange rather than yellow domes. On the sole basis of olfactory stimuli, females showed a significant preference for unripe vs. ripe host fruit, for unripe fruit vs. flowers or leaves, and for host vs. non‐host fruit (or control). However, colour interacted with odour as females dispatched their eggs equally between the yellow dome and the bright orange dome when unripe fruit of tomato was placed under the yellow dome vs. ripe fruit under the bright orange dome. When offered real ripe and unripe tomatoes, females preferred unripe tomatoes. Females significantly chose to lay eggs in non‐infested fruit when they were given the choice between these or fruit infested with larvae. In contrast, recent stings containing eggs did not deter females from laying eggs. Rather, they could have an attractive effect when deposited within <1 h. Regardless of their natal host plant, tomato or bugweed, N. cyanescens females laid significantly more eggs in a dome containing bugweed fruit. However, 15% of females originating from tomato laid eggs exclusively in the dome with tomato, against 3% of females originating from bugweed.     

Intra- and interspecific competition by Fopius arisanus and Diachasmimorpha tryoni (Hymenoptera: Braconidae), parasitoids of tephritid fruit flies

Fopius arisanus (Sonan) and Diachasmimorpha tryoni (Cameron) are two important solitary endoparasitoids of tephritid fruit flies. The former species attacks host eggs while the latter attacks host larvae, and both species emerge as adults from the host puparium. This study investigated intrinsic competition between these two parasitoids, as well as aspects of intraspecific competition within each species in the Mediterranean fruit fly, Ceratitis capitata (Wiedemann). Parasitization by F. arisanus resulted in direct mortality of host eggs and prolonged development of host eggs and larvae. Superparasitism by F. arisanus was uncommon when mean parasitism per host patch was <50%, but increased with rising rates of parasitism. Superparasitism by D. tryoni was more common. In superparasitized hosts, supernumerary individuals of F. arisanus were killed through physiological suppression, while supernumerary larvae of D. tryoni were killed mainly through physical attack. In multiparasitized hosts, dissections showed that 81.6% of D. tryoni eggs in the presence of F. arisanus larvae died within 3 days, indicating physiological inhibition of egg hatch. Rearing results further showed that F. arisanus won almost all competitions against D. tryoni. The ratio of D. tryoni stings to ovipositions was lower in hosts not previously parasitized by F. arisanus than in parasitized hosts, suggesting that D. tryoni can discriminate against parasitized hosts. The mechanism that F. arisanus employs to eliminate D. tryoni is similar to that it uses against all other larval fruit fly parasitoids so far reported. The results are discussed in relation to the competitive superiority of early acting species in fruit fly parasitoids, and to a possible competitive-mediated mechanism underlying host shift by D. tryoni to attack non-target flies following the successful introduction of F. arisanus in Hawaii.     

Clutch size in frugivorous insects as a function of host firmness: the case of the tephritid fly Anastrepha ludens

Abstract. 1. Optimal clutch size theory predicts that individuals will oviposit the number of eggs that increases their fitness. In Anastrepha ludens Loew (Diptera: Tephritidae), females oviposit larger clutches in unripe (firm) fruits than in ripe (soft) fruits. The following hypotheses were tested: (1) Using fruit firmness as an indicator of fruit quality, A. ludens females vary the number of eggs per clutch every time they reach an oviposition decision. (2) Maximising offspring survival with respect to either unripe or ripe fruit requires placing large clutches in firm fruit and smaller clutches in soft fruit. 2. Agar spheres were used as artificial hosts. Three agar concentrations resulted in three degrees of firmness. Mango fruits Mangifera indica L. served as natural hosts. Ripe and unripe fruits were used to test soft and firm host conditions respectively. Females laid significantly larger clutches in the firmer artificial hosts than in the softer hosts. They also laid significantly more eggs in artificial hosts without sugar than in hosts with sugar. Firm (unripe) mangoes also received significantly larger clutches than soft (ripe) mangoes. 3. When an individual female was first presented with a firm artificial host, it laid a large clutch. If subsequently offered a soft host, the female laid a significantly smaller clutch. Finally, if again offered a firm host, clutch size was increased significantly. 4. Possible trade‐offs in offspring fitness were explored in ripe and unripe mangoes by measuring offspring egg‐to‐adult survival, pupal weight, mean adult longevity, and fecundity. Despite the fact that larval survival was greater in soft fruit than in firm fruit, parameters such as pupal weight, mean longevity, and fecundity of adults stemming from both fruit types did not differ significantly. 5. A probable trade‐off between high offspring mortality caused by host unsuitability and low offspring and adult mortality caused by parasitism and predation is discussed as the reason for the exploitation of sub‐optimal hosts.     

Host-marking by female pepper weevils, Anthonomus eugenii

Pepper weevils, Anthonomus eugenii Cano (Coleoptera: Curculionidae), feed and oviposit in flower buds and small fruits of plants in the genus Capsicum, as well as several species of Solanum (Solanaceae). Females chew a small hole into the fruit, deposit a single egg within the cavity, and seal the hole with a clear anal secretion that hardens into an ‘oviposition plug’. Female oviposition behavior was studied in a series of small‐arena bioassays to determine whether previous oviposition in Jalapeño pepper fruit deterred subsequent oviposition and to determine what specific cues from an infested fruit influence female behavior. In choice and no‐choice tests, females preferred clean fruit to fruit that had received four eggs 24 h previously (i.e., infested fruit), whether the fruit was infested with conspecific eggs or their own eggs. Further bioassays demonstrated that the presence of female frass, or oviposition plugs alone, in the absence of eggs or any fruit damage, was sufficient to deter oviposition. In addition, females given the choice between an infested fruit with the oviposition plug removed or an unaltered infested fruit preferred the fruit with no plugs, even when eggs, frass, and feeding damage were still present. To determine whether females would avoid infested peppers under more natural conditions, we quantified oviposition on infested and uninfested sentinel pepper fruit within individually caged plants and on clean and infested plants caged together. Females consistently laid more eggs on clean fruit than on infested fruits and moved within and among pepper plants to search for more acceptable oviposition sites. We conclude that oviposition plugs, along with contaminated female, but not male, frass contain a deterrent that, in the absence of any other cue, is enough to alert a female that a patch is occupied.     

Importance of plant physical cues in host acceptance for oviposition by Busseola fusca

Plant and surrogate stems exhibiting specific combinations of physical cues were used to determine which plant‐related stimuli influence the oviposition of Busseola fusca (Fuller) (Lepidoptera: Noctuidae). The number of eggs and egg batches laid per female increased with an increase in diameter of both natural and artificial stems. Direct observations of the oviposition behaviour (walking, antennating, and sweeping with the ovipositor) indicated that the female moths preferred oviposition supports with a large diameter and non‐pubescent or smooth surfaces over pubescent or rough ones. Pubescence and rough surfaces significantly affected the behavioural steps leading to oviposition by interfering with the ovipositor sweep process necessary to find a suitable oviposition site. Furthermore, more eggs and egg batches were laid on soft than rigid supports. The rigidity of the support affected the proper insertion of the ovipositor for egg deposition. Our results underline the importance of physical stimuli in B. fusca's choice of an oviposition site, which may facilitate the identification of potential host plants or preferred oviposition sites on a plant for this species.     

Biological attributes of three introduced parasitoids as natural enemies of fruit flies,genus Anastrepha (Diptera: Tephritidae)

The biological attributes of three introduced species of parasitoids which attack the fruit fly Anastrepha ludens were evaluated. Larvae and eggs of A. ludens were exposed to larval parasitoids Diachasmimorpha longicaudata and D. tryoni and the egg parasitoid Fopius arisanus. Parasitoid longevity and fecundity were determined using larvae and eggs of A. ludens. Likewise, the parasitism rates of these parasitoid species in infested host fruits were recorded. The intrinsic rate of increase for F. arisanus was 0.1019 followed by D. tryoni with a rate of 0.1641 and D. longicaudata with the highest rate of 0.2233. Although F. arisanus females had the highest levels of fecundity, only 50% of them remained alive until reproductive age. These results in combination with the longer generation time (in comparison with D. longicaudata and D. tryoni), can be considered as the most important factors explaining F. arisanus reduced rate of increase. However, we note that oviposition activity caused egg mortality which reduced Anastrepha egg hatch by ca. 20%. This result suggests that F. arisanus has a high potential as a natural enemy of A. ludens, in accordance with our research efforts to develop a new F. arisanus strain specialized for development in Anastrepha eggs. The results show that D. tryoni is not a good candidate for biological control of Anastrepha. Meanwhile, D. longicaudata continues to be the most important exotic parasitoid for suppression of Anastrepha fruit fly populations.     

CRISPR/Cas9‐mediated mutagenesis of the white gene in the tephritid pest Bactrocera tryoni

The Queensland fruit fly, Bactrocera tryoni (Froggatt), is a polyphagous horticultural pest in Australia that is capable of causing significant damage to more than 100 different host fruits and vegetables. Chemical applications and ecological control strategies, such as the sterile insect technique (SIT), are commonly used to suppress established populations and eradicate invasive outbreaks following migration. The recently published B. tryoni draft genome provides new opportunities to identify candidate genes for targeted genome modification in order to generate advanced genetic strains for management using sterile insect strategies. Here, we demonstrate CRISPR/Cas‐mediated mutagenesis in B. tryoni through generating a series of frame‐shift mutations in the ATP‐dependent binding cassette transporter, white, causing a classic white‐eye phenotype. This work establishes methods for CRISPR/Cas genome editing in tephritids and demonstrates its potential for developing genetic sexing strains which could be used for SIT‐based pest control.     

Effects of pyriproxyfen and buprofezin on immature development and reproduction in the stable fly*

The stable fly, Stomoxys calcitrans (L.) (Diptera: Muscidae), is one of the most significant biting flies that affect cattle. The use of traditional insecticides for stable fly control has only a limited success owing to the insect's unique feeding behaviours and immature development sites. A laboratory study was conducted to evaluate the effects of two insect growth regulator (IGR) products, pyriproxyfen and buprofezin, on the development of the immature stages of the stable fly and the effects of pyriproxyfen on oviposition and egg hatch. Both pyriproxyfen and buprofezin had significant inhibitory effects on immature development. The LC₅₀s of pyriproxyfen and buprofezin were 0.002 and 18.92 p.p.m., respectively. Topical treatment of adult females with different doses of pyriproxyfen had significant negative effects on both female oviposition and egg hatching when 1‐ and 3‐day‐old females were treated, and the effects were dose dependent. A significant reduction in the mean number of eggs laid was observed only at the highest pyriproxyfen dose (8 µg/fly) and egg hatch was unaffected by pyriproxyfen treatment when 5‐day‐old females were treated. Results from the present study indicate that pyriproxyfen has the potential to be used as part of an integrated stable fly management programme.     

Effect of plant oils on quality parameters of Bactrocera tryoni (Froggatt) reared on liquid larval diet

Liquid larval diets have been developed for several tephritid fruit flies including Queensland fruit fly, Bactrocera tryoni (Frogatt) (Q‐fly). In liquid diets, wheat germ oil (WGO) is usually added to improve performance in some quality parameters of reared flies, especially flight ability. However, for some flies, other plant oils may be more readily available, cheaper or produce flies of superior performance. In the present study, four alternative types of plant oils – rice bran, canola, vegetable, and sesame – were incorporated into a fruit fly liquid larval diet to replace the currently used wheat germ oil and their efficacy on the quality parameters of reared Q‐fly was compared to diets containing wheat germ oil or no oil. The quality parameters included: total pupal yield (N), pupal recovery (%), larval duration (days), pupal weight (mg), adult emergence (%), adult fliers (%), rate of fliers (%), sex ratio (%), F₁ egg/female/day and egg hatching (%). There were significant differences among treatments in performance of Q‐fly. Vegetable oil appeared better in terms of total pupal yield, percentage of pupal recovery, percentage of adult emergence, percentage of fliers, mean egg/female/day and % F₁ egg hatch compared with other oil treatments, especially from that of WGO treated diet. The result suggests that WGO can be substituted with rice bran and vegetable oil to improve the liquid larval diet for rearing of B. tryoni, with vegetable oil being the best replacement.