Sawfly Sex Pheromones: Analysis of Their Impact on Pine Odor Attractive to Egg Parasitoids.

Pinus sylvestris trees are known to efficiently defend themselves against eggs of the herbivorous sawfly Diprion pini. Their direct defense against eggs is primable by prior exposure to the sex pheromones of this species and their indirect defense involves attraction of egg parasitoids by egg-induced pine needle odor. But it is unknown whether exposure of pine to D. pini sex pheromones also affects pine indirect defense against sawfly eggs. In this study, we investigated the influence of exposure of P. sylvestris trees to the sex pheromones of D. pini on indirect defense mediated by egg parasitoids. Behavioral assays with Closterocerus ruforum, a key parasitoid of sawfly eggs, revealed no significant attraction to odor from egg-free pines pre-exposed to pheromones. Chemical analyses of odor from egg-free pines showed no pheromone-induced change in the emission rates of the known key terpenoids promoting parasitoid attraction. Further comparative analyses of odor from egg-laden pines pre-exposed to the sex pheromones and of odor from egg-laden pines unexposed to pheromones neither revealed significant differences in the emission rates of terpenoids relevant for parasitoid attraction. The results suggest that a pheromone-induced or pheromone-primed, egg-induced pine indirect defense seems to be redundant in addition to the known pheromone-primable pine direct defense against the eggs and the known egg-inducible indirect defense.

Review of the genus Calotelea Westwood, 1837 (Platygastroidea: Scelionidae) of India.

The genus Calotelea Westwood, 1837 (Scelionidae) of the Indian region is revised. Fifteen new species of Calotelea - C. andamanensis sp. n., C. aurea sp. n., C. brevinotaularis sp. n., C. hodgsoni sp. n., C. kannagiae sp. n., C. lambodara sp. n., C. longistriata sp. n., C. mandavyai sp. n., C. marykingsleyae sp. n., C. microtrichiana sp. n., C. nigriventris sp. n., C. oloftoreni sp. n., C. sibyllamerianae sp. n., C. sushrutai sp. n. and C. trikona sp. n. - are described and imaged. Images of previously described species, C. auriventria Sharma, 1978 and C. immaculata Sharma, 1982, are also provided. Calotelea cameroni (Masner, 1965) is revalidated and redescribed as Probaryconus cameroni Masner, 1965 status revalidated based on morphological characters. Keys to females and known males of Indian species of Calotelea are furnished. While sexual dimorphism with differently coloured males and females has been reported earlier in some species of Calotelea, intrasexual colour dimorphism among females of a species is reported here for the first time.

Behavioural Evidence and Chemical Identification of a Female Sex Pheromone in Anagrus atomus (Hymenoptera: Mymaridae).

Anagrus atomus (L.) is an egg parasitoid involved in the biological control of Empoasca vitis (Göthe) in vineyards. Sex pheromones play a crucial role in mate finding for several parasitoid species and could be used for monitoring under field conditions. We carried out laboratory and field studies aimed at assessing the existence and identity of a possible A. atomus sex pheromone. We found that males were significantly attracted by virgin females independent of age. Males were not attracted to individuals of the same sex, but they were attracted by a crude extract from an unmated female and its polar fraction. Eugenol (4-allyl-2-methoxyphenol) was identified as the attractive substance and proved to be attractive not only in the olfactometer but also in another laboratory bioassay and under field conditions. Attraction of males, but not females, confirms that this is not an aggregation pheromone. This is the first sex-pheromone component identified in Mymaridae, however more compounds could be involved in the mating behaviour of A. atomus. The utility of a sex pheromone in A. atomus is discussed in the context of fitness returns.

Transcriptome sequence-based phylogeny of chalcidoid wasps (Hymenoptera: Chalcidoidea) reveals a history of rapid radiations, convergence, and evolutionary success.

Chalcidoidea are a megadiverse group of mostly parasitoid wasps of major ecological and economical importance that are omnipresent in almost all extant terrestrial habitats. The timing and pattern of chalcidoid diversification is so far poorly understood and has left many important questions on the evolutionary history of Chalcidoidea unanswered. In this study, we infer the early divergence events within Chalcidoidea and address the question of whether or not ancestral chalcidoids were small egg parasitoids. We also trace the evolution of some key traits: jumping ability, development of enlarged hind femora, and associations with figs. Our phylogenetic inference is based on the analysis of 3,239 single-copy genes across 48 chalcidoid wasps and outgroups representatives. We applied an innovative a posteriori evaluation approach to molecular clock-dating based on nine carefully validated fossils, resulting in the first molecular clock-based estimation of deep Chalcidoidea divergence times. Our results suggest a late Jurassic origin of Chalcidoidea, with a first divergence of morphologically and biologically distinct groups in the early to mid Cretaceous, between 129 and 81 million years ago (mya). Diversification of most extant lineages happened rapidly after the Cretaceous in the early Paleogene, between 75 and 53 mya. The inferred Chalcidoidea tree suggests a transition from ancestral minute egg parasitoids to larger-bodied parasitoids of other host stages during the early history of chalcidoid evolution. The ability to jump evolved independently at least three times, namely in Eupelmidae, Encyrtidae, and Tanaostigmatidae. Furthermore, the large-bodied strongly sclerotized species with enlarged hind femora in Chalcididae and Leucospidae are not closely related. Finally, the close association of some chalcidoid wasps with figs, either as pollinators, or as inquilines/gallers or as parasitoids, likely evolved at least twice independently: in the Eocene, giving rise to fig pollinators, and in the Oligocene or Miocene, resulting in non-pollinating fig-wasps, including gallers and parasitoids. The origins of very speciose lineages (e.g., Mymaridae, Eulophidae, Pteromalinae) are evenly spread across the period of chalcidoid evolution from early Cretaceous to the late Eocene. Several shifts in biology and morphology (e.g., in host exploitation, body shape and size, life history), each followed by rapid radiations, have likely enabled the evolutionary success of Chalcidoidea.

Side-effects of pesticides used in irrigated rice areas on Telenomus podisi Ashmead (Hymenoptera: Platygastridae).

Telenomus podisi Ashmead (Hymenoptera: Platygastridae) is an important agent for the biological control of stink bug eggs in irrigated rice areas and the best strategy for its preservation is the use of selective pesticides. The aim of this study was to know the side-effects of pesticides used in Brazilian irrigated rice areas on egg parasitoid T. podisi. We evaluated, under laboratory conditions, 13 insecticides, 11 fungicides, 11 herbicides, and a control (distilled water) in choice and no-choice tests. In the no-choice tests, the pesticides were sprayed at pre and post-parasitism stages (egg and larval stages of T. podisi). In the choice tests, sprays were conducted only at pre-parasitism stages. For all tests, we prepared cards with 25 eggs of the alternative host Euschistus heros (Fabricius) (Hemiptera: Pentatomidae) non-parasitized (pre-parasitism) and parasitized (post-parasitism), which were subjected to pesticide sprays. The parasitism and emergence rates of T. podisi were determined classifying the pesticides in terms of the reduction of parasitism or emergence rates compared to the control. The neurotoxic insecticide cypermethrin, lambda-cyhalothrin, zeta-cypermethrin, etofenprox, thiamethoxam, thiamethoxam + lambda-cyhalothrin, acetamiprid + alpha-cypermethrin, and bifenthrin + alpha-cypermethrin + carbosulfan were more harmful to T. podisi and, therefore, are less suitable for the integrated management of insect pests in irrigated rice areas.

Volatiles Mediating Parasitism of Euschistus conspersus and Halyomorpha halys Eggs by Telenomus podisi and Trissolcus erugatus.

This study identified chemicals found on the eggs of two stink bug species, one native to western North America, Euschistus conspersus, and an invasive species from Asia, Halyomorpha halys. The responses of two scelionid egg parasitoids, Trissolcus erugatus and Telenomus podisi, toward natural stink bug egg volatiles, and synthetic reconstructions of egg volatiles, were tested in bioassays. A compound, methyl (2E,4Z)-2,4-decadienoate, previously identified as the major component of the male-produced aggregation pheromone of E. conspersus, was the major volatile identified from extracts of E. conspersus eggs. In contrast, for H. halys, the sesquiterpenoids that compose the male-produced aggregation pheromone of this species were not detected on eggs, whereas the presence of hexadecanal, octadecanal, and eicosanal was detected. In laboratory olfactometer tests, both Tr. erugatus and Te. podisi females were attracted to extracts of E. conspersus eggs, and to synthetic methyl (2E,4Z)-2,4-decadienoate. However, female Tr. erugatus and Te. podisi wasps were repelled, both by extracts of H. halys eggs and by a blend of the aldehydes identified from H. halys eggs. A follow-up field study, using hexane-washed and intact E. conspersus as sentinel eggs, showed that the parasitoids Trissolcus erugatus and Gryon obesum emerged from these eggs. Sentinel hexane-washed eggs treated with 3 ng of methyl (2E,4Z)-2,4-decadienoate were parasitized more by these two species than were hexane-washed or unwashed eggs, whereas hexane-washed eggs treated with a comparable dose of the C16,18,20 aldehyde mixture were avoided by these parasitoids. In a further field experiment, Trissolcus basalis was the primary parasitoid found in sticky traps baited with methyl (2E,4Z)-2,4-decadienoate, indicating that this species was attracted to, but either did not oviposit or develop in the E. conspersus sentinel eggs in the previous experiment.

Alternating temperatures affect the performance of Trichogramma species.

The environmental compatibility of a biological control agent is an important aspect of successful reduction of agricultural pests. Temperature fluctuations during the day have a strong influence on the performance of laboratory-reared parasitoids. In field conditions, Trichogramma (Hymenoptera: Trichogrammatidae) wasps are exposed to variable temperatures during their development, which has a significant impact on their ability to control pest species. A simulation-based study was undertaken to evaluate the impact of variations in daily temperature on the pest-control abilities of female Trichogramma and their immature progenies. Considering the temperature variability of different agricultural zones of India, five temperature levels ranging from 20ºC to 40ºC were selected for daily short-term heat shocks to the immature progenies and egg-laying females of two major Trichogramma species. Intensity and frequency of thermal shocks showed inverse relationships with adult emergence, fecundity, and longevity of T. chilonis and T. poliae. Parasitoid pupae were found to be more tolerant to temperature variations than eggs and larvae. Fecundity and longevity of parasitoids were significantly reduced under high temperature shocks to egg-laying females. Sex ratio was significantly affected by high temperature shocks to the immature and adult stages. However, the effect was more severe in eggs. A female-biased sex ratio was apparent in both parasitoids throughout the experiment. Overall, daily short-term temperature shocks to different developmental stages of parasitoids showed radical effects on emergence, fecundity, longevity, and sex ratio of the progeny. Therefore, releases of parasitoids should be conducted when they are in their pupal stages during the morning and evening in order to achieve their highest effectiveness for pest management.

First Record of Telenomus dilophonotae (Hymenoptera, Scelionidae), Parasitizing Eggs of Erinnyis ello (Lepidoptera, Sphingidae) in Western Paraná, Brazil, with Molecular Characterization and Records of Occurrences.

There are few records for Telenomus dilophonotae Cameron, 1913 (Hymenoptera, Scelionidae) from South America. In Brazil, the first occurrence was reported in Bahia in rubber crops, Hevea brasiliensis (Willd. ex Adr. de Juss.) Muell. - Arg., there parasitizing eggs of Erinnyis ello Linnaeus, 1758 (Lepidoptera, Sphingidae). It was also found parasitizing the same host in cassava, Manihot esculenta Crantz (Euphorbiaceae). This is the first record of occurrence of T. dilophonotae in the state of Paraná, parasitizing eggs of E. ello in areas of cassava production in the western region of Paraná, this being the southernmost record of the species. Here, photographs, the first sequence of DNA barcode of this species of parasitoid wasp, and a distribution map are provided.

Historical Note on the Genus Trichogramma (Hymenoptera, Trichogrammatidae) in Brazil, Focusing on Taxonomy and Diversity.

The history of the taxonomy of Trichogramma parasitoid wasps can be divided into two phases. In the first phase, species identifications were based on external morphological characters; however, these identifications proved to be confusing and inaccurate. In the second phase, starting in the 1970s, taxa were identified based on charcteristics of the male genital capsule , leading to a major advance in Trichogramma taxonomy. The history of Trichogramma taxonomy in Brazil is recent and mainly related to species that parasitize agricultural pests. In Brazil, the first phase of Trichogramma taxonomy occurred in the 1960s, while the second phase occurred from the 1980s onward. In this second phase, Trichogramma taxonomy progressed significantly and knowledge of Trichogramma diversity as well as associations with lepidopteran pests improved markedly in Brazil as well as worldwide. The last five decades have seen significant progress in studies in Brazil, with taxonomy evolving from identifications based exclusively on morphological characters to integrative taxonomy, encompassing biology (crosses) and morphometry. This historical outline presents the phases of Trichogramma taxonomy in Brazil, addressing the hurdles encountered in the first descriptions, erroneous records of the species, and species descriptions since the 1980s. We highlight the importance of accurately identifying Trichogramma taxa for their use in biological control, as well as species diversity and associations with lepidopteran hosts.

Field Control Effect of Telenomus remus Nixon and Trichogramma chilonis Ishii Compound Parasitoid Balls against Spodoptera frugiperda (J. E. Smith).

Although the release of egg parasitoids has proven to be an effective strategy for the control of the fall armyworm (FAW), a single egg parasitoid, however, has exposed some deficiencies in practice, and it is worthwhile to explore whether the combination of multiple parasitoid species released can be used to control FAW by adopting an inter- or intra-specific relationship. In this study, we released compound parasitoid balls of Te. remus and Tr. chilonis in maize fields to explore the effects of combinations in different proportions of these two egg parasitoids on the control of the FAW. The results showed that the release of compound parasitoid balls improved the control effect on the FAW compared to the release of only Te. Remus (100%) and only Tr. Chilonis (100%). The treatments released with compound parasitoid balls significantly increased the egg parasitism rate against the FAW, reduced its populations, and alleviated its damage to maize as compared to the treatment with only Tr. chilonis (100%) released, whereas there was no significant difference in the egg parasitism rate and FAW populations between the treatments with the release of only Te. remus (100%) and that of compound parasitoid balls. Among the compound parasitoid balls with different proportions of egg parasitoids, the treatment with 80% Te. remus and 20% Tr. chilonis and that with 20% Te. remus and 80% Tr. chilonis could still retain more than 50% of the control effect against the FAW after 15 days of release and given the higher cost of production of Te. Remus, our results suggested that the combination of 20% Te. Remus and 80% Tr. Chilonis is a more cost-efficient ratio for the control of the FAW. Our findings may provide a new perspective for the sustainable control of the FAW.

Review of the genus Sparasion Latreille, 1802 (Hymenoptera: Platygastroidea: Sparasionidae) of the Oriental region with descriptions of new species from India.

BACKGROUND: The genus Sparasion, endoparasitoids of Tettigoniidae, occur in the Nearctic, Palearctic, Afrotropical and Oriental regions. It is absent in the Neotropics and Australasia. Of the thirteen species found in the Oriental region only a single species is from India. RESULTS: Two new species groups - Sparasion bilahari species group and Sparasion manavati species group - are proposed for species from the Oriental region. Thirty-six species are described and illustrated of which twenty-four are new: Sparasion albopilosellus Cameron, 1906 (Pakistan); S. bhairavi Veenakumari, sp. n. (India); S. bhupali Veenakumari, sp. n. (India); S. bihagi Veenakumari, sp. n. (India); S. bilahari Veenakumari, sp. n. (India); S. cellularis Strand, 1913 (Taiwan); S. coconcus Kozlov and Lê, 2000 (Vietnam); S. coeruleus Kieffer, 1905 (Sumatra); S. cullaris Kozlov and Lê, 2000 (Vietnam); S. darbari Veenakumari, sp. n. (India); S. deepaki Veenakumari, sp. n. (India); S. domes Kozlov and Lê, 2000 (Vietnam); S. elbakyanae Veenakumari, sp. n. (India); S. formosus Kieffer, 1910 (Taiwan); S. hindoli Veenakumari, sp. n. (India); S. kalyani Veenakumari, sp. n. (India); S. kanakangi Veenakumari, sp. n. (India); S. karivadana Veenakumari, sp. n. (India); S. lividus Johnson, Masner & Musetti, 2008 (Philippines); S. manavati Veenakumari, sp. n. (India); S. meghmalhari Veenakumari, sp. n. (India); S. micromerus Kozlov and Lê, 2000 (Vietnam); S. pahadi Veenakumari, sp. n. (India); S. philippinensis Kieffer, 1913 (Philippines); S. ratnangi Veenakumari, sp. n. (India); S. rupavati Veenakumari, sp. n. (India); S. salagami Veenakumari, sp. n. (India); S. shulini Veenakumari, sp. n. (India); S. sinensis Walker, 1852 (China); S. sivaranjini Veenakumari, sp. n. (India); S. syamalangi Veenakumari, sp. n. (India); S. todi Veenakumari, sp. n. (India); S. travancoricus Mani and Sharma, 1981 (India); S. vanaspati Veenakumari, sp. n. (India); S. visvambari Veenakumari, sp. n. (India) and S. zeelafi Veenakumari, sp. n. (India). Keys to Oriental species of Sparasion are furnished. Intrasexual colour morphs among females of Sparasion is reported. Lectotype is designated for Sparasion cellularis Strand. CONCLUSIONS: Twenty-four new species are added to the Indian fauna of Sparasion. Previously described species of Oriental Sparasion are redescribed and illustrated. Keys are furnished for all Oriental species.

Factors Affecting Progeny Production and Sex Ratio of Gryon aetherium (Hymenoptera: Scelionidae), a Candidate Biological Control Agent for Bagrada hilaris (Hemiptera: Pentatomidae).

Manipulating the factors that influence progeny production and sex ratio in parasitoids can help maximize the production of quarantine bioassays and/or mass releases. In a series of experiments, we studied the effects of several factors on offspring production and sex ratio in the parasitoid Gryon aetherium (Hymenoptera: Scelionidae), a candidate biological control agent for Bagrada hilaris (Hemiptera: Pentatomidae). Progeny production was influenced by maternal age and dropped when females were 24 or 28 days old and decreased on the second day of exposure. Overall, the offspring sex ratio was highly variable in G. aetherium and was affected by the duration of exposure, with higher proportions of females emerging after one day of exposure than after two days, but was not affected by female density, female age/host deprivation, or temperature during oviposition. Progeny production was affected by the temperature during oviposition and was highest at 26.6 °C. The results indicate that production of G. aetherium can be maximized at one day of exposure, using females that are less than 24 d old, and at temperatures of around 26 °C.

Release strategies of Telenomus podisi for control of Euschistus heros: a computational modeling approach.

BACKGROUND: The egg-parasitoid wasp Telenomus podisi has received attention as a biological-control agent for one of the most important soybean pests in Brazil, the stink bug Euschistus heros. As yet, no studies have conclusively established strategies for the release of T. podisi. We developed a computational model using cellular automata in the C programming language to investigate release strategies for T. podisi in soybean crops, in order to optimize the use of these wasps in managing E. heros, assuming a two-dimensional grid of cells corresponding to a soybean field. RESULTS: The release strategies capable of maintaining an E. heros population below the Economic Threshold level involved releasing a total of at least 15 000 female parasitoids per hectare, in three or four releases of 5000 or more (equivalent to approximately 7142 or more male and female parasitoids per hectare, assuming a sex ratio of 0.70). A 25-m spacing between release points or strips was indicated. The model is very sensitive to the variation in the number of parasitoids per release and in the number of releases, but little sensitive to the release mode and spacing values. CONCLUSION: The theoretical results produced by the computational model are expected to guide future field studies to improve T. podisi release plans for managing E. heros in soybeans. Therefore, we recommend the release strategy of three to four releases of 5000 or more female parasitoids per hectare, at points or strips spaced 25 m apart, to be tested in field experiments for proper implementation by producers. © 2022 Society of Chemical Industry.

Contrasting reproductive traits of competing parasitoids facilitate coexistence on a shared host pest in a biological control perspective.

BACKGROUND: Interspecific competition in insect parasitoids is an important ecological phenomenon that has relevant implications for biological pest control. To date, interspecific intrinsic (=larval) competition has been intensively studied, while investigations on extrinsic (=adult) competition have often lagged behind. In this study we examined the role played by parasitoid reproductive traits and host clutch size on the outcome of extrinsic competition between Trissolcus basalis (Wollaston) and Ooencyrtus telenomicida (Vassiliev), two egg parasitoids of the pest Nezara viridula (L). Laboratory experiments were conducted by allowing both parasitoid species to exploit an egg mass made of 10, 20, 30, or 40 hosts through single or simultaneous releases. Furthermore, under field conditions, egg masses consisting of 10 or 40 hosts were exposed in a tomato crop in order to validate laboratory investigation. RESULTS: The results show that the egg mass size is an important predictor of extrinsic competition in our study system as a higher proportion of T. basalis emerged from large egg masses, while O. telenomicida dominated in small egg masses. Analysis of reproductive traits of parasitoid species indicates that T. basalis has superior abilities in host exploitation compared with O. telenomicida. CONCLUSIONS: We found that contrasting reproductive traits of two competing egg parasitoid species facilitate coexistence on a shared stink bug host. This work also highlights the importance to consider extrinsic competitive interactions between parasitoid species in a biological control perspective. © 2022 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Impact of Environmental Variables on Parasitism and Emergence of Trichogramma pretiosum, Telenomus remus and Telenomus podisi.

Environmental variables may markedly influence egg parasitoid performance and must be considered when choosing the best release strategy. Thus, this study aimed to investigate the effects of light, temperature, soil moisture, and precipitation on the emergence and parasitism of parasitoid releases of unprotected and encapsulated pupae. The presence of light favored the parasitism of Trichogramma pretiosum Riley, 1879, and Telenomus remus (Nixon, 1937) but did not impact the parasitism of Telenomus podisi (Ashmead, 1893). The release strategy adopted (release of encapsulated or unprotected pupae) also impacted the results. On one hand, card capsules gave protection against rainfall, limited to 10 mm regarding Tr. pretiosum, while on the other hand, card capsules led to a reduction in the number of captured adults for Te. podisi at 25°C and 30°C. Therefore, Tr. pretiosum, Te. remus, and Te. podisi can be released using both encapsulated and unprotected pupae, with advantages and disadvantages for each strategy, depending on each studied environmental variable. In addition, parasitoid pupae should be released so that the majority emerge during daylight, especially for Te. remus and Tr. pretiosum, since parasitism was greatly reduced in dark environments. Telenomus podisi is not affected by this variable as it demonstrated similar parasitism in light and dark environments.

Laboratory and field studies supporting augmentation biological control of oriental fruit moth, Grapholita molesta (Lepidoptera: Tortricidae), using Trichogramma dendrolimi (Hymenoptera: Trichogrammatidae).

BACKGROUND: Oriental fruit moth, Grapholita molesta (Busck) (Lepidoptera: Tortricidae) is a cosmopolitan pests of fruit trees in temperate regions. Control of G. molesta is challenging because larvae feed in concealed locations and have evolved resistance to many pesticides. We compared three commercially available species of Trichogramma for efficacy against G. molesta in the laboratory assays and tested releases of the promising species in a pear orchard. RESULTS: Laboratory assays indicated that Trichogramma dendrolimi parasitized G. molesta at the highest rate. Parasitoids took longer to oviposit in older host eggs, and fewer eggs were parasitized when they were more than 3 days old. Field tests produced ca 60% cumulative parasitism of sentinel G. molesta eggs with one release of Trichogramma dendrolimi, with most parasitism occurring within 24 h. Female wasps dispersed up to 12 m from release points with a bias toward upwind movement. CONCLUSION: We calculated that, for each generation of G. molesta, 900 000 wasps per hectare, in three releases 3 days apart, each distributed among release points 10 m apart, would reduce fruit damage by half in an orchard where 50% of fruit would otherwise be damaged. Although augmentation of Trichogramma dendrolimi is a viable tactic for reducing G. molesta populations and fruit damage, it will require integration with other compatible control tactics in order to provide commercially acceptable levels of control in orchards experiencing significant pest pressure. © 2021 Society of Chemical Industry.

An Effective Cold Storage Method for Stockpiling Halyomorpha halys (Hemiptera: Pentatomidae) Eggs for Field Surveys and Laboratory Rearing of Trissolcus japonicus (Hymenoptera: Scelionidae).

An effective stockpiling method for egg masses of the invasive brown marmorated stink bug (Halyomorpha halys [Stål]; Hemiptera: Pentatomidae) would be useful for rearing and field studies of its egg parasitoid Trissolcus japonicus (Ashmead) (Hymenoptera: Scelionidae). The current method of stockpiling H. halys egg masses at subzero temperatures has lethal and sublethal fitness consequences for T. japonicus. Our goal was to test the efficacy of refrigeration as an alternative method for stockpiling H. halys eggs. We show that parasitoid emergence from egg masses refrigerated at 8°C for up to two months before parasitism is higher than from frozen egg masses. In addition, 8°C cold storage usually had minimal or no sublethal fitness effects on emerging T. japonicus. Only after two months of host egg refrigeration did the emergence of T. japonicus begin to decrease significantly (by ~10% relative to untreated viable egg masses), whereas egg masses previously frozen at -80°C had a ~59% reduction in parasitoid emergence after 14 d of storage. Refrigerated egg masses that were subsequently exposed to a range of field-realistic average temperatures were still suitable for T. japonicus parasitism after 7 d. Our results demonstrate that refrigeration at 8°C, while resulting in complete mortality of H. halys embryos after 10 d, are more suitable for T. japonicus parasitism than those stored at -80°C. The quantity and quality of H. halys eggs that can be stockpiled with this method could facilitate T. japonicus laboratory colony maintenance, field monitoring, and releases.

Native and Non-Native Egg Parasitoids Associated with Brown Marmorated Stink Bug (Halyomorpha halys [Stål, 1855]; Hemiptera: Pentatomidae) in Western Slovenia.

Halyomorpha halys (Hemiptera: Pentatomidae), native to East Asia, has become a globally invasive pest, as a serious threat to agricultural production and a notorious nuisance pest in urban areas. Considerable efforts have been made so far to develop effective pest control measures to prevent crop damage. Biological control of this invasive stink bug by egg parasitoids has proven to be the most environmentally sustainable long-term solution. Knowledge of the native egg parasitoid fauna is of key importance when implementing a biological control program. Therefore, the main objective of our study was to detect egg parasitoid species associated with H. halys in the Goriska region (Western Slovenia) and to evaluate their impact on the pest population under field conditions. In the years 2019 and 2020, around 4600 H. halys eggs were collected in the wild and more than 3400 sentinel eggs were exposed to detect parasitoids in the field. Five egg-parasitoid species emerged from H. halys eggs: Anastatus bifasciatus (Hymenoptera: Eupelmidae), Telenomus sp., Trissolcus basalis, Trissolcus mitsukurii (Hymenoptera: Scelionidae) and Ooencyrtus telenomicida (Hymenoptera: Encyrtidae), all of them are new records for Slovenia. The native species, An. bifasciatus, dominated in urban and suburban areas, while non-native Tr. mitsukurii prevailed in agricultural areas. Overall parasitism rates of naturally laid eggs by the parasitoid species complex in 2019 and 2020 was 3.0 and 14.4%, respectively. Rapid recruitment of native parasitoids, early detection of an effective alien parasitoid species and increasing overall parasitism rates are very encouraging results, which need to be followed and verified in future research.

Parasitism and Suitability of Aprostocetus brevipedicellus on Chinese Oak Silkworm, Antheraea pernyi, a Dominant Factitious Host.

Aprostocetus brevipedicellus, a eulophid gregarious egg parasitoid of lepidopterous pests, is a potential biological control agent for the control of many forest pests. A dominant factitious host, Antheraea pernyi, has been widely used for mass rearing several parasitoids in China. However, whether A. pernyi eggs are suitable for A. brevipedicellus rearing remains unclear. Here we evaluated A. brevipedicellus parasitism and fitness of their offspring on A. pernyi eggs with five different treatments, including manually-extracted, unfertilized and washed eggs (MUW), naturally-laid, unfertilized and washed eggs (NUW), naturally-laid, unfertilized, and unwashed (NUUW) eggs, naturally-laid, fertilized and washed eggs (NFW), and naturally-laid, fertilized and unwashed eggs (NFUW). The results showed that A. brevipedicellus could parasitize host eggs in all treatments but significantly preferred MUW eggs to other treatments. Moreover, A. brevipedicellus preferred unfertilized eggs to fertilized eggs and parasitized more washed eggs than unwashed. The pre-emergence time of parasitoid offspring emerging from fertilized eggs was shorter than that from unfertilized eggs. More parasitoid offspring emerged from unwashed eggs than that from washed eggs. The offspring emergence rate was high (>95%) and also female-biased (>85%) among all egg treatments. The egg load of female parasitoid offspring emerging from MUW and NUW eggs was 30-60% higher than the remaining treatments. Overall, MUW eggs of A. pernyi are the most suitable for the mass production of A. brevipedicellus.

The impact of cold storage durations on Trichogramma evanescens (Westwood) (Hymenoptera: Trichogrammatidae) during their pupal stage.

This experimental study was done at the Biological Control Laboratory, Sakha Agricultural Research Station, Sakha, Kafrelsheikh, Egypt. We aimed to estimate the impact of different cold (10 °C) storage durations [0 (non-cold-stored parasitized eggs), 3, 6, 9, 12, 15, 18, 21, 24, 27, and 30 days], on Trichogramma evanescens (Westwood) (Hymenoptera: Trichogrammatidae) during the pupal stage using the eggs of Sitotroga cerealella after exposing to T. evanescens. The emergence percentage of non-cold-stored eggs of S. cerealella was higher than all cold-stored durations. Also, the female's percentages of T. evanescens in the cold storage durations were lower than the non-cold storage one, and they were influenced by extended cold storage durations. There were non-significant differences in the female's longevity of T. evanescens obtained from 0, 3, and 6 days cold-stored parasitized eggs of S. cerealella at 10 °C, but it began to decrease from those produced after 9 days of cold-stored eggs. In addition, the emergence percentage in F1 progeny of T. evanescens was greater than 50% until 21 days of cold storage. It could be concluded that cold storage reduced the % emergence, % females, female's longevity, and emergence percentage in F1 progeny of T. evanescens. For a successful biological control program, the decrease of T. evanescens performance after cold storage durations should be considered in mass production, and the release percentage should be increased by the equivalent of a lack of % emergence. Also, the economic importance of using cold storage periods in commercial mass rearing should be assessed in the biological control program.