The egg parasitoid Anastatus japonicus is a key natural enemy in the biological control of various agricultural and forestry pests. It is particularly used against the brown marmorated stink bug Halyomorpha halys and the emerging defoliator pest Caligula japonica in East Asia. It has been proved that the eggs of Antheraea pernyi can be used as a factitious host for the mass production of A. japonicus. This study systematically documented the parasitic behaviour and developmental morphology exhibited by A. japonicus on the eggs of A. pernyi. The parasitic behaviour of A. japonicus encompassed ten steps including searching, antennation, locating, digging, probing, detecting, oviposition, host-feeding, grooming, and resting. Oviposition, in particular, was observed to occur in three stages, with the parasitoids releasing eggs during the second stage when the body remained relatively static. Among all the steps of parasitic behaviour, probing accounted for the longest time, constituting 33.1% of the whole time. It was followed by digging (19.3%), oviposition (18.5%), antennation (9.6%), detecting (7.4%), and the remaining steps, each occupying less than 5.0% of the total event time. The pre-emergence of adult A. japonicus involves four stages: egg (0 to 2nd day), larva (3rd to 9th day), pre-pupa (10th to 13th day), pupa (14th to 22nd day), and subsequent development into an adult. Typically, it takes 25.60 ± 0.30 days to develop from an egg to an adult at 25℃. This information increases the understanding of the biology of A. japonicus and may provide a reference for optimising reproductive devices.

The egg parasitoid Cleruchoides noackae Lin & Huber, 2007 (Hymenoptera: Mymaridae) is originated from Australia and the main biological control agent of Thaumastocoris peregrinus Carpenter & Dellapé, 2006 (Hemiptera: Thaumastocoridae) on Eucalyptus L'Hér (Myrtaceae). Companies that grow Eucalyptus are in need of a mass rearing protocol to increase the number of individuals produced and improve the quality of this parasitoid. The aim of this study was to define a protocol for mass rearing C. noackae in T. peregrinus eggs, based in the evaluations of the key biological attributes of this parasitoid in the parental and F1 generations, after the cold storage of the parasitised host eggs. Two methods were tested as C. noackae rearing protocols. In the first, parasitised eggs of T. peregrinus by C. noackae were cold stored for 7 days after being left in a climatic chamber at 24 ± 2°C, 60 ± 10% RH and a photoperiod of 12:12 (light:dark) h (standard environmental conditions) for 3, 6, 9 or 12 days. In the second, T. peregrinus eggs parasitised by C. noackae were maintained in a climatic chamber under standard environmental conditions for 6 days, after which these eggs were cold-stored for 0 (control), 7, 14 or 21 days. Parasitism (%), and the development period (parasitism to adult) and female proportion (%) of C. noackae were evaluated. Based on the results (parental generation: parasitism, around 45%; F1 generation: parasitism, around 55%; development period, around 16 days; female proportion, around 60%), eggs should be stored at 5°C on the sixth day after parasitism by C. noackae and maintained at this temperature for 7 days. The cold storage of T. peregrinus eggs, after parasitism, can be included in the mass rearing protocols of the parasitoid C. noackae.

Ooencyrtus pityocampae and Ooencyrtus kuvanae are egg parasitoids that are considered potential candidates for the control of different pest species through inundative release. The aim of this study was to assess the effects of different cold-storage periods of Philosamia ricini eggs (host) on the rearing parameters of O. pityocampae and O. kuvanae. Host eggs were stored at 3 °C, and a factorial experiment involving two parasitoid species, nine host storage periods (1, 5, 10, 15, 30, 45, 60, 75 and 90 days) and a control, and two host ages (1 and 2 days) was conducted, with 10 replications including 40-P. ricini eggs each. Adult emergence, development time, longevity, and fecundity were investigated. The parasitoid adult emergence percentage significantly varied with storage duration. These values were lower in O. kuvanae than in O. pityocampae. The development time of O. kuvanae progeny increased in both host age groups except in the 1-day storage period subgroup. However, the development times of the progeny of O. pityocampae reared on one-day-old eggs stored for 5, 10, 60, and 75 days were increased, and the development times of the progeny of O. pityocampae reared on 2-day-old eggs stored for 45 and 90 days were increased. The longevity of the F1 progeny of O. kuvanae was negatively affected by storage time. There was no difference in the longevity of the F1 progeny of O. pityocampae compared to that of the control. Additionally, the fecundities of the F1 progeny of O. pityocampae and O. kuvanae were 54.7 and 47.0 offspring/female, respectively. These results provide useful information for guiding the development of mass rearing methodologies for both parasitoid species.

Plants not only respond to herbivorous damage but adjust their defense system after egg deposition by pest insects. Thereby, parasitoids use oviposition-induced plant volatiles to locate their hosts. We investigated the olfactory behavioral responses of Trichogramma pretiosum Riley, 1879 (Hymenoptera: Trichogrammatidae) to volatile blends emitted by maize (Zea mays L.) with singular and stacked events after oviposition by Spodoptera frugiperda Smith, 1797 (Hymenoptera: Trichogrammatidae) moths. Additionally, we examined possible variations in gene expression and on oviposition-induced volatiles. We used a Y-tube olfactometer to test for the wasp responses to volatiles released by maize plants oviposited by S. frugiperda and not-oviposited plants. Using the real-time PCR technique (qRT-PCR), we analyzed the expression of lipoxygenase and three terpene synthases genes, which are enzymes involved in the synthesis of volatile compounds that attract parasitoids of S. frugiperda. Olfactometer tests showed that T. pretiosum is strongly attracted by volatiles from transgenic maize emitted by S. frugiperda oviposition (VTPRO 3, more than 75% individuals were attracted). The relative expression of genes TPS10, LOX e STC was higher in transgenic hybrids than in the conventional (isogenic line) hybrids. The GC-MS analysis revealed that some volatile compounds are released exclusively by transgenic maize. This study provides evidence that transgenic hybrids enhanced chemical cues under oviposition-induction and helped to increase T. pretiosum efficiency in S. frugiperda control. This finding shows that among the evaluated hybrids, genetically modified hybrids can improve the biological control programs, since they potentialize the egg parasitoid foraging, integrating pest management.

Insecticide application and augmentative parasitoid releases are often considered incompatible. However, pesticide applications and parasitoid releases can be integrated into a pest management scheme if there is careful time scheduling of these interventions. In this study, we assessed the influence of commonly used insecticides (chlorpyrifos-methyl, deltamethrin, pyriproxyfen, thiamethoxam) in olive agroecosystems to two currently present Trichogramma parasitoids in the Mediterranean basin. Exposure to insecticides in relation to parasitoid's development was also tested. Both, insecticide type and application time influenced parasitism and the emergence rates of the two parasitoid species. Chlorpyrifos-methyl had the strongest impact on parasitoids resulting in low numbers of emerged adults followed by deltamethrin. The two parasitoids also exhibited different levels of susceptibility to the insecticides used. Potential integration of insecticides to integrated pest management using Trichogramma parasitoids is discussed.

Egg parasitoid rearing on factitious hosts is an important step to reduce costs and increase availability of the biocontrol agent but it requires quality control to achieve success in field conditions. To this end, this study evaluated the quality of Telenomus remus (Hymenoptera: Platygastridae) reared on Corcyra cephalonica (Lepidoptera: Pyralidae) for until 45 generations. In the first bioassay, we evaluated the body size of the laboratory-produced parasitoids. In the second bioassay, flight activity was examined, measuring the percentage of ‘flyers’, ‘walkers’ and ‘deformed’ parasitoids. The third bioassay assessed parasitism on Spodoptera frugiperda (Lepidoptera: Noctuidae) eggs. Our data indicate that the laboratory-reared parasitoid neither lost its ability to fly nor to parasitize S. frugiperda eggs. In conclusion, quality did not decrease significantly during 45 generations, and therefore rearing of T. remus on C. cephalonica as factitious host promises to be successful.

The egg parasitoid Trissolcus vassilievi (Mayr) (Hymenoptera: Scelionidae) is a significant natural enemy of the sunn pest, Eurygaster integriceps Puton (Hemiptera: Scutelleridae), the most important pest of wheat in Iran. This study examined the developmental time and egg-to-adult survival of two geographically separate populations of T. vassilievi on two corresponding host populations at five constant temperatures ranging from 15.0 to 35.0±1°C. No wasps of either population emerged at 15.0°C and the temperature threshold for development was similar between populations, ranging from 13.1±0.3 to 13.8±0.4°C for males and 12.2±0.1 to 12.6±0.1°C for females, but the thermal constant varied with gender and parasitoid population. Development of wasps from the colder Tabriz location was slower, with thermal constants for males and females of 172.6±3.1 and 204.1±1.2 degree-days, respectively, compared to Varamin wasps with 164.7±3.0 and 195.6±1.3 degree-days, respectively. Based on genetic inheritance patterns, reciprocal crosses between the two populations were expected to result in females with thermal phenotypes intermediate to their parental populations, and males that resembled their mothers. However, female progeny of crosses more closely resembled their maternal population, indicating a maternal effect on thermal phenotype. Furthermore, the magnitude of the maternal effect on the thermal constant was asymmetric and was more strongly expressed by Varmin than Tabriz females. These results suggest the possibility of using selective crosses between wasp populations, in combination with artificial selection in the laboratory, to tune the thermal phenotype of parasitoids to specific regions prior to augmentative releases.

Genetically modified crops with insect resistance genes from Bacillus thuringiensis Berliner (Bt-plants) are increasingly being cultivated worldwide. Therefore, it is critical to improve our knowledge of their direct or indirect impact not only on target pests but also on non-target arthropods. Hence, this study evaluates comparative leaf consumption and performance of Spodoptera eridania (Cramer), a species that is tolerant of the Cry1Ac protein, fed with Bt soybean, MON 87701×MON 89788 or its non-Bt isoline. We also assessed the comparative performance of the egg parasitoid Telenomus remus Nixon on eggs of S. eridania produced from individuals that fed on these two soybean isolines as larvae. Results showed that Bt soybean reduced by 2 days larval development and increased by 3 days adult male longevity. Therefore, we conclude that the effect of Bt soybean MON 87701×MON 89788 on S. eridania development and reproduction is small, and favorable to pest development. These differences are less likely to directly result from the toxin presence but indirectly from unintended changes in plant characteristics caused by the insertion of the transgene. Our results should be viewed as an alert that S. eridania populations may increase in Bt soybeans, but on the other hand, no adverse effects of this technology were observed for the egg parasitoid T. remus which can help to prevent S. eridania outbreaks on these crops.

The egg parasitoid Telenomus busseolae Gahan is the most important parasitoid of sugarcane pink borers and plays an important role in the suppression of their populations in Iran. To understand the efficiency of T. busseolae parasitizing Sesamia cretica Lederer, in this study, the functional response of the parasitoid was investigated at different temperatures (20, 25, 30 and 35 °C). The results of logistic regression indicated a type III functional response at all the temperatures. Roger's random parasite equation fitted the observed data at 30 and 35 °C, but Holling's disc equation fitted the data better at 20 and 25 °C. The attack rate of T. busseolae ranged from 0.0036 to 0.2148, 0.00017 to 0.1031, 0.0049 to 0.2952 and 0.0006 to 0.0382 per h, and the estimated handling time was 0.80, 0.48, 0.32 and 0.13 h at 20, 25, 30 and 35 °C, respectively.

In order to succeed in biological control programs, not only is it crucial to understand the number of natural enemies to be released but also on how many sites per area this releasing must be performed. These variables might differ deeply among egg parasitoid species and crops worked. Therefore, these trials were carried out to evaluate the parasitism (%) in eggs of Anticarsia gemmatalis and Pseudoplusia includens after the release of different densities of the egg parasitoid Trichogramma pretiosum. Field dispersal was also studied, in order to determine appropriate recommendations for the release of this parasitoid in soybean fields. The regression analysis between parasitism (%) and densities of the parasitoid indicated a quadratic effect for both A. gemmatalis and P. includens. The maximum parasitism within 24 h after the release was reached with densities of 25.6 and 51.2 parasitoids per host egg, respectively, for the two pests. Parasitism of T. pretiosum in eggs of P. includens decreased linearly as the distance of the pest eggs from the parasitoid release sites increased. For P. includens, the mean radius of T. pretiosum action and the area of parasitoid dispersal in the soybean crop were 8.01 m and 85.18 m2, respectively. We conclude that for a successful biological control program of lepidopteran pests using T. pretiosum in soybean fields, a density of 25.6 parasitoids per host egg, divided into 117 sites per hectare, should be used.

When Telenomus nawai Ashmead or Trichogramma ostriniae Pang & Chen (Hymenoptera: Trichogrammatidae) was inoculated into intact egg masses of Spodoptera litura (Fabricius) that were covered with a scale-hair layer, T. nawai emerged from 76% of the eggs, while emergence of T. ostriniae was from only 3% of the eggs. When the hair layer was removed before inoculation, the rate by the latter increased to 21%. These observations confirmed that the hair layer effectively protects S. litura egg masses from attack by T. ostriniae, and that T. nawai parasitism is more effective. In order to assess the feasibility of T. nawai as a biological control agent for S. litura, parasitism rate, development time and adult longevity were examined in the laboratory. Emergence of T. nawai was observed in more than 95% of 0- and 1-day-old separated eggs of S. litura, but the rate decreased to 60% and 0% for 2- and 3-day-old eggs, respectively. The emergence rates of T. nawai were near 95% for temperatures in a range from 25°C to 35°C, but decreased to 80% at 20°C. No parasitoids emerged at 15°C. The developmental periods decreased as temperature increased from 20°C to 35°C. The developmental threshold occurred at 13.7°C and 13.9°C, and the effective accumulative temperatures from egg to adult emergence were 149.3 and 147.1 degree-days for females and males, respectively. Mean longevity of the adult females decreased with increasing temperature; 87.0 days at 15°C and 9.5 days at 35°C. The feasibility of the use of T. nawai for controlling S. litura is discussed.