Comparison of the thermal performance between a population of the olive fruit fly and its co-adapted parasitoids

Long-term separation of a host from its native parasitoids may result in divergent thermal adaptation between host and parasitoid. The olive fruit fly, Bactrocera oleae (Rossi), most likely originated from Sub-Saharan Africa, but has since had a long invasion history in cultivated olives that spans geographical barriers and continents. This study compared three major thermal performance profiles (development, survival, and reproduction) across a wide range of temperatures (10–34 °C) among a Californian population of the olive fruit fly and two African parasitoids, Psyttalia lounsburyi (Silvestri) and Psyttalia humilis (Silvestri), believed to have co-adapted with the fruit fly in its native range. Temperature ranges for the development and survival were 10–30 °C for the fly, 10–28 °C for P. lounsburyi, and 14–32 °C for P. humilis. There was no difference in any thermal performance measured between two P. humilis populations (Kenya and Namibia) tested. The most suitable temperature ranges for reproduction were 22–30 °C for the fly, 18–32 °C for P. humilis, and 18–26 °C for P. lounsburyi. The results showed slight differences in the thermal profiles among olive fruit fly and both parasitoids species, with P. humilis being more heat tolerant whereas P. lounsburyi was less heat tolerant than the fruit fly. The results are discussed with respect to thermal co-adaptation and classical biological control of the olive fruit fly.     

Proteomic interactions between the parasitoid Diachasmimorpha longicaudata and the oriental fruit fly,Bactrocera dorsalis during host parasitism

The oriental fruit fly, Bactrocera dorsalis, is an important agricultural pest and biological control is one of the most effective control methodologies. We conducted an investigation on the molecular response of the fruit fly to parasitism by the larval parasitoid, Diachasmimorpha longicaudata using two-dimensional gel electrophoresis and mass spectroscopy. We identified 285 differentially expressed protein spots (109 proteins) during parasitism. The molecular processes affected by parasitism varied at different time point during development. Transferrin and muscle specific protein 20 are the only two proteins differentially expressed that play a role in host immunity 24 h after parasitism. Developmental and metabolic proteins from parasitoids (transferrin and enolase) were up-regulated to ensure establishment and early development of parasitoids 48 h post parasitism. 72 h after parasitism, larval cuticle proteins, transferrin and CREG1 were overexpressed to support the survival of parasitoids while host metabolism proteins and parasitoid regulatory proteins were down-regulated. Host development slowed down while parasitoid development went up at 96 h after parasitism. All developmental, regulatory, structural, and metabolic proteins were expressed at their optimum at 120 h post parasitism. Host development was reduced, metabolism and regulatory proteins were strongly involved in the activities. The development deteriorated further at 144 h after parasitism. Enolase and CREG1 were indicators of parasitoid survival. Hexamerin and transferrin from the parasitoid was peaked at 168–216 h after parasitism, strongly indicating that parasitoid would survive. This study represents the first report that reveals the molecular players involved in the interaction between the host and parasitoid.     

Aggressive Behavior in Olive Fruit Fly Females: Oviposition Site Guarding against Parasitic Wasps

Among Tephritidae flies, the females display agonistic behavior to maintain single oviposition sites and reduce larval competition for food. In the olive fruit fly, Bactrocera oleae, female-female aggressive interactions are characterized by reciprocal wing waving, chasing, head butting and boxing with forelegs. Little is known on tephritid aggressive behaviors directed towards natural enemies, with special reference to parasitoids attacking their young instars. In this study, we quantified the aggressive behavior of B. oleae females guarding their oviposition site against the braconid parasitoid Psyttalia concolor. The fly aggressive behavior displayed against the parasitoids was compared that directed towards paper dummies mimicking P. concolor adults. When a P. concolor female came close (<20 mm) to a B. oleae female guarding the oviposition site, the 91.67% of the flies displayed wing-waving, the 63.34% chased the parasitoid, the 45% showed head-butting, while boxing was observed only in the 26.67% of the aggressions. When paper dummies were tested, only the 66.67% of the flies displayed wing-waving, the 8.33% performed chasing, followed by head butting (5.00%) and boxing (3.33%). B. oleae displayed longer aggression bouts towards live wasps over dummies. Overall, this is the first evidence pointing out that tephritid aggressive acts, besides their role in intraspecific interactions, were also highly effective to displace parasitic wasps from the fly oviposition site. Further research on potential consequences on fitness traits arising from the above-discussed behaviors, as well as on parasitoid learning-mediated responses to tephritid aggressions, is urgently required.     

Olive fruit fly adult response to attract-and-kill bait stations in greenhouse cages with weathered bait spray and a commercial table olive orchard

Attract-and-kill bait stations and olive foliage sprayed with insecticidal bait spray and exposed to 0–4 weeks of weather were evaluated for efficacy by olive fruit fly, Bactrocera oleae (Rossi), adult mortality. Mortality in greenhouse cage tests was significantly higher after three days than one day of exposure to non-weathered bait spray on bait stations and foliage, and on bait stations exposed to weather for three weeks. Mortality increased with an increase in insect exposure time, and decreased as bait spray weather exposure increased. Mortality was higher on bait stations than on olive foliage, and treated foliage showed little toxicity after 4 weeks in weather. In a commercial orchard, bait station efficacy was determined by captures of adults on yellow panel traps. Day temperatures were slightly higher on the underside of the station than top from 2 May to 10 July, similar on the top and underside 11 July to 7 August, and slightly lower on the underside from 8 August to 4 September. Fruit length increased from 0.4 on 16 May to 3 cm on 24 September and on 30 May attained 1 cm, the minimum size to produce one adult. No larvae or adults emerged from collected fruit. More adults were captured in an untreated row than a row with bait stations until 22 August and significantly so for the period ending on 11 July. Maximum adult captures occurred during 2 weeks prior to 19 September. Attract-and-kill bait stations would help reduce insecticidal applications and table olive production costs.     

Sugar concentration and timing of feeding affect feeding characteristics and survival of a parasitic wasp

The availability of food sources is important for parasitoid survival, especially for those that inhabit ecosystems where nectar and honeydew are spatially or temporally scarce. Therefore, the value of even a single meal can be crucial for survival. Psyttalia lounsburyi is a parasitoid, and biological control agent, of the olive fruit fly, Bactrocera oleae. In order to improve our understanding of the basic nutritional ecology of P. lounsburyi and its role in survival we evaluated the effect of a single sucrose meal on the longevity of female and male wasps. We measured the duration of feeding, volume ingested, sucrose consumption, energy content, and longevity of wasps provided with different concentrations of sucrose (0.5, 1, and 2 M) at different times after emergence (0, 1, 2 or 3 days after emergence). Our results showed that longevity was significantly influenced by sucrose concentration and timing of feeding. For females, feeding on sucrose increased the likelihood of survival to varying degrees, ranging from 32.3% to 95.4%, compared to water-only controls. The longest duration of feeding was observed for the highest sucrose concentrations and oldest wasps. The amount of sugar ingested and energy uptake increased, up to a point, as sugar concentration increased. Our results suggest that P. lounsburyi derived greatest benefit from the intermediate concentration (1 M) of sucrose provided 2 or 3 days after emergence. Our study emphasizes the importance of finding balance between increasing longevity and limiting the duration of feeding, and concomitant uptake of nutrients, that is fundamental for survival of the wasp in nature.     

Oviposition response and development of the egg-pupal parasitoid Fopius arisanus on Bactrocera oleae, a tephritid fruit fly pest of olive in the Mediterranean basin

To date, information is wanting with regard to the use of new exotic parasitoids against olive fruit fly, Bactrocera (=Dacus) oleae (Gmelin) (Diptera: Tephritidae), a serious pest of olives Olea europaea L., in the Mediterranean basin. We investigated the oviposition response and developmental biology on B. oleae of Fopius (=Biosteres) arisanus (Sonan) (=Opius oophilus Fullaway) (Hymenoptera: Braconidae), an egg-pupal parasitoid of tephritid fruit flies, never tested before as a potential parasitoid of this host. Our results showed that olive fruits infested with B. oleae eggs exerted a relevant attraction to gravid F. arisanus and represented a stimulus for oviposition. Nevertheless they were not as attractive to female parasitoids as the Mediterranean fruit fly, Ceratitis capitata Wiedemann (Diptera: Tephritidae), eggs infested papaya fruits (Carica papaya L.). In our experimental conditions, F. arisanus completed development in B. oleae within 33 ± 1.7 days (males) and 35 ± 1.6 (females). Increases in host egg to female parasitoid ratios of 1:1, 5:1, 10:1 and 20:1 corresponded with decreases in the percentage of B. oleae parasitisation and host killing but corresponded also with increases in absolute parasitisation. Our findings are discussed in light of possibilities of utilising F. arisanus for biological control of olive fruit fly.     

Oviposition and reproductive performance of a generalist parasitoid (Trichogramma pretiosum) exposed to host species that differ in their physical characteristics

The response of generalist egg parasitoids to alternative natural hosts that are present simultaneously is not well known. We investigated the behavior of Trichogramma pretiosum Riley (Hymenoptera: Trichogrammatidae) in relation to two field hosts Helicoverpa armigera Hübner and Spodoptera litura Fabricius, in choice and no choice tests. We quantified the effects of natal host species and post-emergence adult age on the oviposition preference of the parasitoids. H. armigera eggs were consistently preferred over S. litura eggs, regardless of the natal host and adult age. When only S. litura eggs were available as hosts, they were parasitized at statistically similar rates to H. armigera eggs (average of 17 ± 2.7 vs. 13 ± 3.0, H. armigera to S. litura). The adult lifespan and lifetime fecundity of T. pretiosum were variable but were affected by natal host species and/or host species to which they were exposed. Mean lifespan and fecundity of parasitoids that had developed in H. armigera eggs and were exposed to H. armigera eggs for oviposition were 13.9 ± 1.8 days and 98.7 ± 11.0 adult offspring. By contrast, those that developed in S. litura eggs and were exposed to S. litura eggs for oviposition lived for 7 ± 0.9 days and produced 53.8 ± 8.0 adult offspring. The ovigeny index (OI) was significantly lower in the parasitoids exposed to H. armigera eggs than in those exposed to S. litura eggs, regardless of the natal host, indicating that H. armigera eggs sustain the adult parasitoids better than S. litura eggs. These results are used to predict parasitoid behavior in the field when both hosts are available.     

Comparative evaluation of two olive fruit fly parasitoids under varying abiotic conditions

Psyttalia lounsburyi (Silvestri) and P. humilis (Silvestri) (Hymenoptera: Braconidae) were evaluated in California for their potential to control the invasive olive fruit fly, Bactrocera oleae (Rossi) (Diptera: Tephritidae). Psyttalia lounsburyi is a specialist on B. oleae while P. humilis also attacks other tephritid species. Field cage trials, conducted from 2006 to 2009, were used to compare P. lounsburyi and two populations of P. humilis (Kenya and Namibia) in California’s interior valley and coastal regions. Both parasitoid species reproduced on B. oleae in all trials. Under similar abiotic conditions, offspring production per female was higher in P. humilis than in P. lounsburyi, suggesting that host specificity by P. lounsburyi does not confer a higher efficiency on B. oleae in cultivated olives. Two abiotic factors were shown to impact parasitoid efficiency. First, adult parasitoid survival was poor during periods of high summer temperatures, common to the olive production areas in California’s interior valleys. Second, parasitism levels were lower on B. oleae larvae feeding in larger Ascolano cv. fruit than in smaller Manzanillo cv. fruit. Results are discussed relative to biological control of B. oleae in commercial olives and the usefulness of natural enemies specialized to attack fruit flies in wild olives compared with the larger cultivated olive fruit.     

Host suitability and fitness-related parameters of Campoletis sonorensis (Hymenoptera: Ichneumonidae) as a parasitoid of the cabbage looper,Trichoplusia ni (Lepidoptera: Noctuidae)

The seven age-classes of Trichoplusia ni (Hübner) larvae evaluated in this study as hosts of Campoletis sonorensis indicates that early 2nd larval instar (3–5 day-old larvae) of T. ni represents the most suitable host stage for the development of the larval endoparasitoid C. sonorensis. The higher suitability of early 2nd larval instar of T. ni resulted in more parasitised larvae, a higher rate of successful parasitoid emergence, a higher rate of female progeny, and a lower rate of immature parasitoid mortality. The fitness gain of C. sonorensis on late 1st larval instar (2 day-old larvae) and late 2nd larval instar –early 3rd instars (6–8 day-old larvae) stages of T. ni is negatively affected by the trade-offs between the different physiological and behavioral characteristics influencing their suitability as hosts of C. sonorensis.     

Mass‐rearing optimization of the parasitoid Psyttalia lounsburyi for biological control of the olive fruit fly

The olive fruit fly, Bactrocera oleae (Tephritidae), is a direct pest of olives that has invaded the Mediterranean Region and California. Psyttalia lounsburyi (Braconidae), a larval parasitoid from Africa, has been approved for release in the USA as a classical biological agent. However, it has been difficult to rear the parasitoid in the laboratory because it is multivoltine, and the host develops only in fresh olives, which are not available for most of the year. A method to rear the parasitoid on the factitious host, Mediterranean fruit fly (Ceratitis capitata) was developed, but it was not very efficient for producing large numbers of parasitoids needed for release. We developed a number of ways to improve the efficiency of rearing, including the frequency and duration of exposure for oviposition, optimizing the density of adult parasitoids, host age, as well as methods to quickly standardize the number of larvae exposed and to count emerging adult parasitoids. We significantly improved the number of progeny produced per female and the sex ratio of progeny. Thanks to these improvements, we produced in 2017 over 119,000 adults and shipped over 53,900 for release in California.     

Effects of daily fluctuating temperatures on the Drosophila–Leptopilina boulardi parasitoid association

Koinobiont parasitoid insects, which maintain intimate and long-term relationships with their arthropod hosts, constitute an association of ectothermic organisms that is particularly sensitive to temperature variations. Because temperature shows pronounced natural daily fluctuations, we examined if experiments based on a constant temperature range can mask the real effects of the thermal regime on host-parasitoid interactions. The effects of two fluctuating thermal regimes on several developmental parameters of the Drosophila larval parasitoid Leptopilina boulardi were analyzed in this study. Regime 1 included a range of 16–23–16 °C and regime 2 included a range of 16–21–26–21–16 °C (mean temperature 20.1 °C) compared to a 20.1 °C constant temperature. Under an average temperature of 20.1 °C, which corresponds to a cold condition of L. boulardi development, we showed that the success of parasitism is significantly higher under a fluctuating temperature regime than at constant temperature. A fluctuating regime also correlated with a reduced development time of the parasitoids. In contrast, the thermal regime did not affect the ability of Drosophila to resist parasitoid infestation. Finally, we demonstrated that daily temperature fluctuation prevented the entry into diapause for this species, which is normally observed at a constant temperature of 21 °C. Overall, the results reveal that constant temperature experiments can produce misleading results, highlighting the need to study the thermal biology of organisms under fluctuating regimes that reflect natural conditions as closely as possible. This is particularly a major issue in host-parasitoid associations, which constitute a good model to understand the effect of climate warming on interacting species.     

Fruit nutritional composition and non-nutritive traits of indigenous South African tree species

Frugivorous animals play a major role in dispersing tropical, and to a lesser extent, temperate tree species. In order to attract potential seed dispersers, plants generally offer a reward of fleshy fruit pulp. Criteria for fruit choice by avian frugivores are influenced by a number of non-nutritive (e.g. fruit size and colour) factors; and nutritional composition of the fruit. There is a paucity of nutritional composition and other fruit trait data of indigenous South African fruit. This information is necessary in order to determine which frugivores are likely to ingest which fruits and consequently act as potential seed dispersal agents. This information would provide us with an understanding of the inter-relationships between indigenous fruit and frugivores in South Africa. Consequently nutritional composition was investigated in various indigenous fruit species that avian frugivores feed on. Fruits were collected from 38 indigenous tree species found in KwaZulu-Natal Afromontane and coastal forests. Pulp was freeze-dried to constant mass and then analysed for sugar, lipid and protein content; and for water content determination. Fruit width in this study ranged from 4 mm (Searsia rehmanniana and Trema orientalis) to 40 mm (Annona senegalensis, Ficus sur and Xylotheca kraussiana). Of the fruits examined in this study 29% were black and 43% were red when ripe. Most (84%) fruit species analysed for sugar content were hexose dominant with 50% being fructose and 34% being glucose dominant. Only 16% of the fruit species analysed were sucrose dominant. Fruits in this study were generally observed to be high (mean: 68.1 ± 3.3%; n = 30) in water content; and low in protein and lipid content respectively (mean: 8.2 ± 0.5%; 9.3 ± 2.2%; n = 30) indicating that these fruit species could be considered as nutrient-dilute. Future studies need to determine the nutritional composition of the remaining indigenous South African fruit in order to develop a comprehensive database as well as examining non-nutritive factors.     

Effects of a lipid-rich diet on adult parasitoid income resources and survival

Effectiveness of parasitoids as natural enemies in agro-ecosystems depends on key traits such as fecundity and longevity. Energy sources allocated into survival and reproduction can be mobilized from capital stores acquired during larval feeding, or from income resources through adult feeding. Adult parasitoids have a restricted diet consisting solely of carbohydrates and most species do not replenish lipid levels after emergence from their host. Here, we have adopted a novel approach that could improve pest elimination by parasitoids, in which we added different ratios of olive oil to the food of adults to reveal the potential of a lipid-rich diet to increase lipid levels and allocation into survival. Our results show that males of the parasitoid Cotesia glomerata had higher lipid levels when fed a diet containing a ratio of 90:10% honey to olive oil when compared to males at emergence and males fed honey-only. For females, lipid levels at emergence were similar to those of females fed a diet supplemented with a ratio of 75:25% honey to olive oil, yet in both of these treatments lipid levels were significantly higher when compared to females fed honey-only. This suggests that females on a lipid-containing diet economized on their lipid use. In contrast, the survival of males and females was negatively affected by the addition of olive oil and no differences in survival were found when wasps were fed 1 day on various ratios of honey to olive oil compared to honey-only. Our results show that the addition of a lipid source can increase or maintain nutrient availability and further research into lipid supplementation could minimize detrimental effects in order to increase the effectiveness of parasitoids in certain agro-ecosystems.     

Ruminal degradability,digestibility, energy content,and influence on nitrogen turnover of various Mediterranean by-products in fat-tailed Awassi sheep

Livestock husbandry in the dry areas of the Mediterranean basin is facing scarcity and fluctuation of feed supply and feed prices. At the same time the local agro-food industry is expanding and its by-products might be used as alternatives to traditional feed resources, but their nutritional value is not well known. Therefore, four by-products typical for the Mediterranean area were tested in vitro (n = 3 per diet or feed), in sacco (n = 6) and in vivo (n = 6) either individually or in a proportion of 0.34 of the total diet. The test feeds (broken lentils, sugar beet pulp, tomato pomace and crude olive cake) were compared to a barley–wheat bran mixture (control). Forage (barley straw) made up proportionately 0.5 of the diet. For all experiments either rumen canulated or intact castrated male Awassi sheep were employed. The thirty sheep used in the in vivo experiment weighed on average 42 kg. The animals were fed 1.1 kg dry matter (DM)/day and had free access to water. Large compositional differences (g/kg DM) between the batches of test feeds used in the present study were found in crude protein (from 79 to 245; for olive cake and broken lentils), neutral detergent fiber (aNDFom; from 283 to 584; for broken lentils and olive cake) and non-fiber carbohydrates (from 109 to 436 for tomato pomace and olive cake and to broken lentils). Two feeds (tomato pomace and olive cake) were rich in ether extract; sugar beet pulp was rich in calcium. Broken lentils had the highest effective degradabilities (ED). Olive cake was lowest in ED of OM and aNDFom (0.39 and 0.32, respectively). Olive cake caused refusals of concentrate (lower proportionate concentrate intake compared with the control, P<0.001). The apparent nutrient digestibility of broken lentils and sugar beet pulp was similar (P<0.001) to the control and lower (P<0.001) for tomato pomace and olive cake. Metabolizable energy (ME; MJ/kg DM) was determined by various ways and always showed a very low value for olive cake (1–2). Broken lentils and sugar beet pulp had ME contents higher (P<0.001) than that of tomato pomace. The study showed that some by-products were promising as alternatives to traditional concentrate ingredients, while others, especially olive cake, might be only used at low levels.     

Short-range cues mediate parasitoid searching behavior on maize: The role of oviposition-induced plant synomones

Host searching by egg parasitoids faces a main constraint due to low detectability of cues from host eggs. Therefore egg parasitoids have developed distinctive strategies by exploiting cues that originate from non-target instars of the host and/or from plants. The scelionid Telenomus busseolae is specialized on concealed eggs of Sesamia nonagrioides and other noctuid stemborers. In this paper we show that oviposition by S. nonagrioides induces changes in the cues present on maize leaf surface, which arrest naïve females of T. busseolae. The induction appears to be systemic as the parasitoid also responds to leaves and leaf portions that are not directly affected by ovipositing females. Such oviposition-induced, short-range, plant synomones, acting in sequence with the kairomonal cues from scales left on the plant by the ovipositing host female, significantly increases parasitoid efficiency during host finding. The elicitor of plant response originates from the host female reproductive system, being contained both in the host’s ovarian eggs and in the colleterial gland secretion. Induction starts 24 h after oviposition and lasts at least till 72 h. The ecological role of this oviposition-induced plant synomone in host searching by T. busseolae is discussed.     

Potential risks of systemic imidacloprid to parasitoid natural enemies of a cerambycid attacking Eucalyptus

A community of insect herbivores has established on eucalyptus species in California following their introduction from Australia. A number of the species are under complete or partial biological control. A response to introduction of additional pest species into the complex has been the application of systemic insecticides to infested trees. Natural enemies that have been introduced to control the various pest species feed on the nectar of treated trees, and thus may be affected by these pesticides. In this study, Eucalyptus rudis trees were treated at label rates with the neonicotinoid systemic insecticide imidacloprid and nectar sampled at 5 months post-treatment during the spring bloom. The concentration of imidacloprid and its toxic metabolites in nectar was measured by ELISA at 660 ppb. Adults of the encyrtid egg parasitoid Avetianella longoi that were fed floral nectar collected from treated trees had significantly lower survival and reproductive fitness than adults fed nectar from untreated trees. In feeding bioassays, in which the adults were fed a range of concentrations of imidacloprid in sugar water, the LC₅₀ for A. longoi was 212 ppb imidacloprid. Bioassays were also conducted with the braconid larval parasitoid, Syngaster lepidus. The LC₅₀ for S. lepidus was 288 ppb imidacloprid. The insecticide appears to be concentrating in the nectar at higher levels than reported from other plant species and at concentrations exceeding the LC₅₀ for two important parasitoids. If tree treatments become widespread as a result of continual introductions of new eucalypt herbivores, established biological control programs could be at significant risk.     

Interactions between Phaedrotoma scabriventris Nixon (Hymenoptera: Braconidae) and Diglyphus isaea Walker (Hymenoptera: Eulophidae), parasitoids of Liriomyza huidobrensis (Blanchard) (Diptera: Agromyzidae)

Liriomyza leafminer flies represent a serious threat to horticultural production in East Africa. Total field parasitism rates recorded in Kenya are below 5%, with the indigenous ectoparasitoid Diglyphus isaea Walker being one of the key parasitoid species. The International Centre of Insect Physiology and Ecology (icipe), in collaboration with the International Potato Centre (CIP), imported into Kenya the endoparasitoid Phaedrotoma scabriventris Nixon to improve natural control of leafminers. The objective of this study was to investigate the interactions between D. isaea and P. scabriventris when used together for the biological control of Liriomyza species. These interactions were studied under laboratory conditions, using treatments that involved single, simultaneous and sequential releases of the different parasitoid species onto plants infested by L. huidobrensis larvae. While used separately, parasitism rates of D. isaea and P. scabriventris were 30.4 ± 10.9% and 63.6 ± 7.7% respectively. However, when used simultaneously, the total parasitism rate increased to 77.0 ± 5.3%. Although P. scabriventris had no effect on D. isaea, the presence of D. isaea reduced the specific parasitism rate of P. scabriventris. In addition, both parasitoids induced leafminer mortality through larval-feeding and stinging. However, feeding and stinging mortality induced by D. isaea (41.9 ± 9.1%) was significantly higher compared to P. scabriventris (11.9 ± 8.7). Similarly, pupal mortality due to feeding and stinging activity was 49.1 ± 6.5% and 21.6 ± 1.9% when exposed to D. isaea and P. scabriventris respectively. The implication for simultaneous use of both parasitoids in East Africa is discussed.     

ITS2 sequence variations among members of Diachasmimorpha longicaudata complex (Hymenoptera: Braconidae) in Thailand

The parasitoid Diachasmimorpha longicaudata complex in Thailand contains at least 3 cryptic species informally designated as species D. longicaudata A, B and BB. DNA sequence data of nuclear ITS2 (second internal transcribed spacer) were used to characterize members of this D. longicaudata complex. The polymerase chain reaction (PCR) amplicon of ITS2 region of D. longicaudata B (≈ 650 bp) clearly differentiated this species from A and BB (amplicon of ≈ 590 bp). Sequence alignment of individual parasitoids revealed that low intraspecies differences ranged from 0.457 to 3.991%, but interspecies differences ranged from 7.566 to 12.989%. Phylogenetic trees constructed using Neighbor-Joining (NJ) and Maximum Parsimony (MP) methods, taking the parasitoid Psyttalia concolor complex as an outgroup, revealed that D. longicaudata A, B and BB formed a monophyletic group, with species A and BB being more closely related than species B. ITS2 characterization of D. longicaudata complex has revealed an interesting divergence of the three cryptic sibling species in Thailand.     

Performance of diapausing parasitoid wasps,Habrobracon hebetor,after cold storage

The ectoparasitoid Habrobracon hebetor (Say) (Hymenoptera: Braconidae) is an important potential biological control agent for lepidopterous pests of stored products. We investigated the effects of long-term cold storage of diapausing and nondiapausing H. hebetor on their performance after cold storage. Mortality during storage increased with increasing storage duration, and the mortality of diapausing females was lower than that of nondiapausing females after 8, 12, and 16 weeks of storage. Longevity, egg laying, number of progeny produced, and time to 50% egg laying were all reduced, as compared with the culture females when parasitoids were reared at conditions that do not induce diapause. But, for females reared at 20 °C at conditions that induce diapause, all of these quality parameters did not differ from those of culture insects when the storage duration was 8 weeks or less. The percentage of female F1 offspring was always lower for cold stored insects than for the culture insects. Presence of a male after cold storage did not impact any of the quality parameters measured. Thus, rearing parasitoids at 20 °C and 10L:14D and then storing them for up to 8 weeks at 5 °C would produce parasitoids that are similar to culture parasitoids, except that the percentage of females is lower than that in the cultures (36% vs. 52%).