Biocontrol Programs Research Articles

Impact of diet and bacterial supplementation regimes on Orius strigicollis microbiota and life history performance

Given the growing interest in manipulating microbiota to enhance the fitness of mass-reared insects for biological control, this study investigated the impact of an artificial diet on the microbiota composition and performance of Orius strigicollis. We compared the microbiota of O. strigicollis fed on an artificial diet and moth eggs via culturing and 16S rRNA gene amplicon sequencing. Subsequently, we assessed life history traits and immune gene expression of O. strigicollis fed on the artificial diet supplemented with Pantoea dispersa OS1. Results showed that microbial diversity remained largely unaffected by the artificial diet, with similar microbiota compositions in both diet groups. OS1, a minor member of the microbiota but significantly enriched in bugs fed on the artificial diet, improved nymphal survival rates and shifted adult longevity-reproduction life history in females. Additionally, OS1 supplementation elevated the transcription of antimicrobial peptide diptericin. According to population parameters, the group receiving OS1 only during the nymphal stage showed higher population growth potential compared to the group supplemented across all life stages. These findings reveal the resilience of O. strigicollis microbiota under distinct dietary conditions and highlight the potential of using natural symbionts and specific supplementation regimes to improve Orius rearing for future biocontrol programs.

Reproductive parameters of a new biocontrol agent, Eupeodes americanus (Diptera: Syrphidae) and comparison with the commercialized Aphidoletes aphidimyza (Diptera: Cecidomyiidae).

The American hoverfly Eupeodes americanus (Wiedemann) (Diptera: Syrphidae) is an aphidophagous predator during its larval stage and is currently being evaluated for inclusion in biocontrol programs as a new biocontrol agent. However, little is known about its reproductive aptitudes. The objective of the present study was to determine the reproductive parameters of E. americanus and to compare them with those of a commercialized and widely used biological control agent for aphids, the aphid midge Aphidoletes aphidimyza (Rondani) (Diptera: Cecidomyiidae). The preoviposition period, oviposition period, adult longevity, lifetime and daily fecundity, egg hatching rate, and fertility were determined for E. americanus females and compared to those of A. aphidimyza. Trials were conducted under laboratory conditions in rearing cages on the broad bean plant Vicia faba L. (Fabaceae), infested with pea aphids Acyrthosiphon pisum (Harris) (Hemiptera: Aphididae). The results revealed that the preoviposition period, oviposition period, and adult longevity were significantly longer in E. americanus than in A. aphidimyza. The daily fecundity and egg-hatching rate were similar in both species. However, lifetime fecundity and fertility were considerably higher in E. americanus than in A. aphidimyza. This study demonstrates that the reproductive capacity of E. americanus is clearly superior to that of A. aphidimyza and therefore supports its inclusion in the aphid pest management program as a new biocontrol agent.

Rich diversity of RNA viruses in the biological control agent, Orius laevigatus

Orius laevigatus (Hemiptera, Anthocoridae) is a generalist predator extensively used for the biocontrol of diverse agricultural pests. Previous studies on O. laevigatus have focused on the improvement of insect genetic traits, but little is known about its association with microbes, especially viruses that may influence its production and efficacy. More than 280 RNA viruses have been described in other Hemiptera insects, in line with the continuous discovery of insect-specific viruses (ISVs) boosted by next-generation sequencing. In this study, we characterized the repertoire of RNA viruses associated with O. laevigatus. Its virome comprises 27 RNA viruses, classified within fourteen viral families, of which twenty-three viruses are specific to O. laevigatus and four are likely associated with fungal microbiota. The analysis of viral abundance in five O. laevigatus populations confirmed the presence of simultaneous viral infections and highlighted the ubiquitous presence and high abundance of one solinvivirus and three totiviruses. Moreover, we identified 24 non-retroviral endogenous viral elements (nrEVEs) in the genome of O. laevigatus, suggesting a long-term relationship between the host and its virome. Although no symptoms were described in the insect populations under study, the high diversity of viral species and the high abundance of certain RNA viruses identified indicate that RNA viruses may be significant for the applicability and efficacy of O. laevigatus in biocontrol programs.

Supplementary sugars enhance the production efficiency and parasitism performance of the egg parasitoid Trichogramma dendrolimi (Hymenoptera: Trichogrammatidae).

Trichogramma spp. wasps are egg parasitoids with a long history of mass rearing for augmentation biocontrol programs in field crop and orchard landscapes. Supplementary nutrition can improve the longevity, fecundity, and biocontrol efficacy of parasitoids. To improve the production efficiency and parasitism performance of Trichogramma dendrolimi Matsumura (Hymenoptera: Trichogrammatidae), the present study screened and examined the potential supplementary nutrients for this biological control agent. Dietary supplementation with a 10% sucrose solution significantly increased wasp longevity and parasitism potential of T. dendrolimi on host eggs, but provision of pollen did not provide additional benefits. Laboratory and greenhouse cage tests demonstrated that wasp access to soybean aphid Aphis glycines Matsumura (Hemiptera: Aphididae) honeydew, comprised primarily of melezitose and trehalose, improved T. dendrolimi longevity and parasitism. In conclusion, provision of a 10% sucrose solution to adult wasps will enhance the mass-rearing efficiency of T. dendrolimi; furthermore, field release of T. dendrolimi by plant vectors bearing honeydew-producing aphids holds promise for improving the biocontrol efficacy of T. dendrolimi.

Functional response and predation rate of Cryptolaemus montrouzieri (Coleoptera: Coccinellidae) to Paracoccus marginatus (Hemiptera: Pseudococcidae) at different temperatures.

The ladybug, Cryptolaemus montrouzieri (Mulsant) (Coleoptera: Cocccinellidae)(Mulsant)(Coleoptera: Cocccinellidae), is a highly efficient predator in controlling mealybug populations and is considered an effective agent for controlling the papaya mealybugs (Paracoccus marginatus) (Williams & Granara de Willink) (Hemiptera: Pseudococcidae). Various criteria have been proposed for evaluating predator effectiveness, with the consumption rate of prey by individual predators, specifically the functional response, emerging as a common and crucial metric. This study evaluated the functional responses of third- and fourth-instar larvae, as well as male and female adults (<48 h old) of C. montrouzieri to adult females of P. marginatus at 3 different temperatures (22 °C, 28 °C, and 35 °C) with 70% ± 5% RH and a 12L:12D h photoperiod. Prey densities were 2, 4, 8, 16, 32, 45, or 60 papaya mealybugs per predator for all tests. The response to prey density by third- and fourth-instar larvae or both sexes of adult C. montrouzieri was a type II at all temperatures. The highest attack rate and lowest handling time were estimated at 28 °C in males and 35 °C in females, respectively. The highest daily prey consumption rate occurred at 35 °C in both the immature and adult stages of C. montrouzieri. These findings support the potential of C. montrouzieri in controlling the papaya mealybug, especially in tropical and subtropical regions, given its search efficiency at high temperatures tested in this study. However, additional field investigations are needed to ascertain the control efficacy of C. montrouzieri for this mealybug in biocontrol programs.

Complex nitrogen sources from agro-industrial byproducts: impact on production, multi-stress tolerance, virulence, and quality of Beauveria bassiana blastospores.

Biological control through entomopathogenic fungi provides essential ecological services in the integrated management of agricultural pests. In the context of submerged liquid fermentation, the nutritional composition significantly influences the ecological fitness, virulence and quality of these fungi. This study specifically explores the impact of various complex organic nitrogen sources derived from agro-industrial byproducts on the submerged liquid fermentation of Beauveria bassiana, a versatile entomopathogenic fungus known for producing hydrophilic yeast-like blastospores. Notably, manipulating the nitrogen source during submerged cultivation can influence the quality, fitness, and performance of blastospores. This research identifies cottonseed flour as the optimal low-cost nitrogen source, contributing to increased production yields, enhanced multi-stress tolerance, heightened virulence with extended shelf life and long-term bioactivity. These findings deepen our understanding of the critical role of nitrogen compound selection in liquid media formulation, facilitating the production of ecologically fit and virulent blastospores for more effective pest biocontrol programs.

Genetic matching and the identification of a promising biocontrol agent validates a decision to survey natural enemies of Urena lobata in Malaysia

Urena lobata is a major introduced pasture weed in Vanuatu where a biocontrol program to mitigate its impacts commenced in 2018. There was considerable uncertainty regarding the native range of U. lobata, although published literature, coupled with very simple climate matching suggested that Southeast Asia should be the most promising region in which to survey for natural enemies. We, therefore, conducted surveys for candidate biocontrol agents in Malaysia and conducted genetic matching to compare plants growing in Vanuatu, Malaysia, and other regions in the invaded and purportedly native range of U. lobata. Surveys in Malaysia prioritized a tingid bug Haedus vicarius as a promising candidate agent for the biocontrol of U. lobata and subsequent host specificity testing confirmed it is sufficiently host specific to be released in Vanuatu. Genetic matching indicated that plants growing in Malaysia are a good match to plants present in Vanuatu, validating our selection of survey sites.

Effects of cold storage temperature, host status, and larval stage on the biological characteristics of Cotesia chilonis

AbstractShelf life is an important determinant of the quality of parasitoids used as natural enemies in augmentation biological control. Cotesia chilonis (Munakata) (Hymenoptera: Braconidae), an obligate endoparasitoid of Chilo suppressalis (Walker) (Lepidoptera: Crambidae), has potential for use through augmentation biological control in China. Here, the effects of cold storage temperature, host status, and larval stage on the biological characteristics of C. chilonis were investigated to extend the agent's shelf life. Results showed that the number of healthy C. chilonis adults obtained was reduced by 3.5% and 47.7% when C. suppressalis larvae in either natural or induced diapause were used as hosts compared to nondiapausing control larvae. Both parasitism and cold storage significantly reduced the supercooling point of nondiapausing C. suppressalis larvae, whereas the opposite result was observed in those of natural diapause. In order to improve the long‐term storage of C. chilonis, nondiapausing C. suppressalis larvae were parasitized for 1, 3, 5, and 7 days to allow C. chilonis to develop into different larval stages before being transferred into cold storage for 30 days at 4 or 10°C. Compared to the control, the C. chilonis cocoon formation and emergence rates of the 1‐day‐old egg treatment group held at 10°C were reduced by 15% and 17%, respectively. The results also showed that the duration of the larval stage of C. chilonis was prolonged fourfold after storage at 4 and 10°C. Individuals held at 10°C had a significantly longer cocoon and adult stage compared to those held at 4°C. The biological parameters and life history of the F1 generation of C. chilonis returned to normal levels under laboratory conditions. In conclusion, C. suppressalis larvae were successfully used to extend the shelf life of C. chilonis for use in augmentative biocontrol programs.

Patch exploitation behaviour by a parasitoid of a forest pest: The influence of host cues and intra‐ and interspecific interactions

Abstract For parasitoids, host foraging is the most significant activity for their reproductive success, as it directly translates into offspring production. Therefore, foraging behaviour and patch‐use decisions by parasitoids still attract research interest, especially when the studied species are biological control agents. In laboratory conditions, we studied the patch exploitation behaviour of Megarhyssa nortoni (Hymenoptera: Ichneumonidae), a larval–pupal parasitoid of several woodwasps (Hymenoptera: Siricidae), exposed to intra‐ and interspecific interactions. It is known that M. nortoni females are attracted to the symbiotic fungus carried by its hosts, which support host larval development inside the wood. Additionally, in the introduced range where the species has been released in biocontrol programs aimed at managing invasive Sirex noctilio populations, Megarhyssa nortoni coexists with the egg–larval parasitoid Ibalia leucospoides. First, we investigated whether host–fungal symbiont cues and direct presence of conspecific females affected patch time allocation and oviposition behaviour of M. nortoni, as both stimuli could provide information on concealed hosts. Second, we studied the influence of prior parasitism by I. leucospoides on patch time allocation decisions and oviposition behaviour. The main results showed that females modulated their responses to the fungal cues when foraging simultaneously with conspecifics. Also, patch‐leaving tendencies and patch residence times were not affected when females foraged patches previously exploited by I. leucospoides; indeed, they probed these patches more frequently. Our study contributes to the understanding of patch‐use decisions made by parasitoids and their impact on the biological control of damaging insect populations, particularly in a context of two parasitoid species and one host.

Artificial Adult Diet as a New Tool for Improving a Biocontrol Program with Predatory Hoverflies

Syrphine hoverflies (Diptera: Syrphinae) are important predators of aphids in agricultural crops. While the use of flowering plants to enhance their efficacy is well established, recent research has developed an artificial diet for adult hoverflies consisting of a sugar solution and pollen in a dispenser. To ensure that the artificial diet is suitable to support hoverfly reproduction, a comparative analysis was conducted between a natural diet of flowering buckwheat plants versus an artificial diet consisting of artificial flowers (including honey solution and pollen), complemented by a sugar solution disperser. The study evaluated the fecundity, fertility, oviposition period, egg hatchability, and overall lifespan of the American hoverfly, Eupeodes americanus (Wiedemann 1830). The results indicate that the artificial diet does not negatively impact the reproductive parameters of E. americanus when compared to the buckwheat-based diet. Consequently, artificial diets emerge as a promising and more convenient alternative to flowering plants to support hoverflies in biological control strategies and for their mass rearing in research facilities and commercial insectaries.

Analyzing the safety of the parasiticide fungus Mucor circinelloides: first insights on its virulence profile and interactions with the avian gut microbial community.

Parasiticide fungi are considered an accurate, sustainable, and safe solution for the biocontrol of animal gastrointestinal (GI) parasites. This research provides an initial characterization of the virulence of the native parasiticide fungus Mucor circinelloides (FMV-FR1) and an assessment of its impact on birds' gut microbes. The genome of this fungus was sequenced to identify the genes coding for virulence factors. Also, this fungus was checked for the phenotypic expression of proteinase, lecithinase, DNase, gelatinase, hemolysin, and biofilm production. Finally, an in vivo trial was developed based on feeding M. circinelloides spores to laying hens and peacocks three times a week. Bird feces were collected for 3 months, with total genomic DNA being extracted and subjected to long-read 16S and 25S-28S sequencing. Genes coding for an iron permease (FTR1), iron receptors (FOB1 and FOB2), ADP-ribosylation factors (ARFs) (ARF2 and ARF6), and a GTPase (CDC42) were identified in this M. circinelloides genome. Also, this fungus was positive only for lecithinase activity. The field trial revealed a fecal microbiome dominated by Firmicutes and Proteobacteria in laying hens, and Firmicutes and Bacteroidetes in peacocks, whereas the fecal mycobiome of both bird species was mainly composed of Ascomycetes and Basidiomycetes fungi. Bacterial and fungal alpha-diversities did not differ between sampling time points after M. circinelloides administrations (P = 0.62 and P = 0.15, respectively). Although findings from this research suggest the lack of virulence of this M. circinelloides parasiticide isolate, more complementary in vitro and in vivo research is needed to conclude about the safety of its administration to birds, aiming at controlling their GI parasites.IMPORTANCEA previous study revealed that the native Mucor circinelloides isolate (FMV-FR1) can develop parasiticide activity toward coccidia oocysts, one of the most pathogenic GI parasites in birds. However, ensuring its safety for birds is of utmost importance, namely by studying its virulence profile and potential effect on commensal gut microbes. This initial study revealed that although this M. circinelloides isolate had genes coding for four types of virulence factors-iron permease, iron receptors, ADP-ribosylation factors, and GTPase-and only expressed phenotypically the enzyme lecithinase, the administration of its spores to laying hens and peacocks did not interfere with the abundances and diversities of their gut commensal bacteria and fungi. Although overall results suggest the lack of virulence of this M. circinelloides isolate, more complementary research is needed to conclude about the safety of its administration to birds in the scope of parasite biocontrol programs.

Importance of pathobiomes to the success of microbial weed biocontrol agents

The role of microbial communities in the establishment of plant pathogens introduced to new environments as weed biocontrol agents is relatively unexplored. Substantial evidence from crop protection research shows how bulk microbial communities influence the establishment, survival, and success of pathogens. Just as mismatches between biocontrol agents, host genotypes, and climate can lead to establishment failure, we propose that mismatches between microbial biocontrol agents and resident microbiomes may be a cause of apparent weed biocontrol failures. From this perspective, we discuss recent progress in research on plant-associated microbial communities whose interactions influence disease development, or “pathobiomes,” and consider the implications for biocontrol agent evaluation and release. Open questions about how weed pathobiomes develop and influence agent success rates provide ample opportunity for researchers to adapt new methods and analytical frameworks that will enhance biocontrol programs. We conclude by prioritizing the development of standard methods to include microbiomes as a component of release studies. Developing a pathobiome-aware agent selection and release process will improve agent establishment rates and biocontrol outcomes.

Soil fertility as a mediator of interactions between an introduced specialist beetle and a native generalist nematode on an exotic invasive plant and its native congener

Abstract Invasive plants are often attacked by both introduced specialist and native generalist natural enemies in new ranges. Soil fertility can potentially alter the interactions of these natural enemies with native versus invasive plants in ways that have largely unexplored implications for biological invasions and biological control. A common garden experiment was conducted to compare the performance of an introduced specialist flea beetle, Agasicles hygrophila, and/or a native generalist nematode, Meloidogyne incognita, on invasive alligator weed, Alternanthera philoxeroides, and its native congener sessile joyweed, Alternanthera sessilis, under different levels of soil nitrogen (N) and phosphorus (P). At a relatively low or moderate N and P levels, the flea beetle and the nematodes were not significantly affected by each other. Under these conditions, alligator weed responded plastically by producing more branches and biomass, as well as longer stolons, in response to attack by the flea beetle and/or the nematode, compared to sessile joyweed responses to similar levels of damage. However, under a relatively high N and P levels, nematode infestations significantly reduced flea beetle damage on alligator weed, resulting in significantly greater above‐ and below‐ground biomass and longer stolons than plants without herbivory. In contrast, beetle herbivory significantly increased the level of nematode infestations on sessile joyweed, resulting in significantly fewer fine and coarse roots, and lower above‐ and below‐ground biomass compared to plants without herbivory. Synthesis and applications. Our findings illustrate the importance of soil fertility in mediating interactions between specialist biocontrol agents and native generalists on native versus invasive plants. High phenotypic plasticity seems to be an important attribute that contributes to the success of invasive plants like alligator weed in relatively nutrient‐poor environments. However, nutrient‐rich environments could potentially confer greater growth benefits on invasive plants than on native ones by changing herbivore–herbivore interactions on plants differently. There is a great need to fully investigate the direct and indirect interactions between biocontrol agents and generalists across food webs following classical biocontrol releases. Nutrient measurements of both soil and water bodies should also be incorporated into all stages of biocontrol programmes.

Manipulated ants: inducing loyalty to sugar feeders with an alkaloid.

Wood ants are promising biocontrol agents in fruit plantations because they prey on pest insects and inhibit plant diseases. However, these ants also attend plant-feeding homopterans to harvest their honeydew secretions, thereby increasing their numbers. This problem can be solved by offering ants alternative sugar sources that are more attractive than honeydew. From natural interactions, it is known that some species manipulate mutualistic partners toward loyalty by adding alkaloids to the food they offer their mutualists. Inspired by this, the addition of alkaloids might be used to make ants loyal to artificial sugar feeders and thus used to reduce populations of ant-farmed homopterans in ant-mediated biological control. We aimed to explore whether wood ants (Formica polyctena) would develop a taste preference for morphine-containing sugar solutions in two-choice laboratory tests. After having fed on a morphine/sugar solution for 1 week, ants showed a significant preference for morphine solutions compared with equal concentration sugar solutions without morphine. Furthermore, ants lost this preference after 6-9 days on a morphine-free diet. The results show that wood ants react to morphine in their food, enabling chemical manipulation of their behavior, most likely through a taste preference. Thus, ants are susceptible to manipulation by mutualistic partners in natural interactions and furthermore may be manipulated artificially in biocontrol programs to avoid ant-mediated build-up of homopteran populations. © 2024 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Surveillance and screening of Stomoxyinae flies from Mallorca Island (Spain) reveal the absence of selected pathogens but confirm the presence of the endosymbiotic bacterium Wolbachia pipientis

Adult brachycera biting flies can significantly impact livestock through both direct effects (reduction of food intake, disturbance, painful bites, and blood loss) and indirect effects (pathogen transmission), leading to substantial economic losses and production damage. This study aimed to assess the presence of blood-sucking flies in six mixed-animal farm environments on the island of Mallorca (Balearic Islands, Spain) by employing multiple trapping methods. Additionally, distribution maps of brachycera biting fly species recorded in Spain were created, based on data extracted thorough review of scientific literature and citizen digital databases. Investigation of several pathogens, including equine infectious anemia virus (EIAV), Anaplasmataceae bacteria, and piroplasm protozoa, was carried out using different PCR targets (18S rRNA, 16S rRNA, groESL, and tat genes). Citizen science databases and literature review corroborated the consistent distribution trend for two Stomoxyinae species, underscoring the importance of citizen collaboration as a complement to traditional entomological surveillance. Our study confirmed the presence of two biting Stomoxyinae species: the prevalent stable fly Stomoxys calcitrans across all sampled farms, and the horn fly Haematobia irritans, which turned out to be less abundant. DNA barcoding techniques validated the identification of the two species. Neither EIAV nor bacterial/protozoan pathogens were detected using the selected PCR targets in either fly species. However, Wolbachia pipientis (clustered in the supergroup A together with the only sequence of W. pipientis from the USA) was identified through PCR targeting 16S rRNA, groESL and wsp genes in all pools of H. irritans (n = 13) collected from two of the examined farms. This study represents the first attempt to investigate pathogens in Stomoxyinae biting flies in Spain. The discovery of the endosymbiotic Wolbachia organism in H. irritans represents the first record in Spain and the second from Europe. This finding holds significant implications for future research on the applications of this bacterium in biocontrol programs.

The Diversity of Parasitoids and Their Role in the Control of the Siberian Moth, Dendrolimus sibiricus (Lepidoptera: Lasiocampidae), a Major Coniferous Pest in Northern Asia.

The Siberian moth, Dendrolimus sibiricus Tschetv., 1908 (Lepidoptera: Lasiocampidae) is a conifer pest that causes unprecedented forest mortality in Northern Asia, leading to enormous ecological and economic losses. This is the first study summarizing data on the parasitoid diversity and parasitism of this pest over the last 118 years (1905-2022). Based on 860 specimens of freshly reared and archival parasitoids, 16 species from two orders (Hymenoptera and Diptera) were identified morphologically and/or with the use of DNA barcoding. For all of them, data on distribution and hosts and images of parasitoid adults are provided. Among them, the braconid species, Meteorus versicolor (Wesmael, 1835), was documented as a parasitoid of D. sibiricus for the first time. The eastern Palaearctic form, Aleiodes esenbeckii (Hartig, 1838) dendrolimi (Matsumura, 1926), status nov., was resurrected from synonymy as a valid subspecies, and a key for its differentiation from the western Palaearctic subspecies Aleiodes esenbeckii ssp. esenbecki is provided. DNA barcodes of 11 parasitoid species from Siberia, i.e., nine hymenopterans and two dipterans, represented novel records and can be used for accurate molecular genetic identification of species. An exhaustive checklist of parasitoids accounting for 93 species associated with D. sibirisus in northern Asia was compiled. Finally, the literature and original data on parasitism in D. sibiricus populations for the last 83 years (1940-2022) were analysed taking into account the pest population dynamics (i.e., growth, outbreak, decline, and depression phases). A gradual time-lagged increase in egg and pupal parasitism in D. sibiricus populations was detected, with a peak in the pest decline phase. According to long-term observations, the following species are able to cause significant mortality of D. sibiricus in Northern Asia: the hymenopteran egg parasitoids Telenomus tetratomus and Ooencyrtus pinicolus; the larval parasitoids Aleiodes esenbeckii sp. dendrolimi, Cotesia spp., and Glyptapanteles liparidis; and the dipteran pupal parasitoids Masicera sphingivora, Tachina sp., and Blepharipa sp. Their potential should be further explored in order to develop biocontrol programs for this important forest pest.

Direct and indirect host‐related volatile compounds attract a fruit fly parasitoid, Diachasmimorpha longicaudata

AbstractFruit fly pests (Diptera: Tephritidae) are a serious problem for fruit production and for local and international trade. Biological control is increasingly included as a pest control tool within integrated pest management (IPM) programmes, seeking to reduce pesticides and improve fruit quality. Ceratitis capitata Wiedemann, the Mediterranean fruit fly (medfly), is probably the most damaging fruit fly pest, with a global distribution and more than 200 host species. The solitary larval endoparasitoid Diachasmimorpha longicaudata Ashmead (Hymenoptera: Braconidae) is a biocontrol agent widely used against Tephritidae fruit fly pests. Previous studies showed that female wasps locate host larvae using visual, mechanical, and chemical cues. Here, we investigated the chemical basis of female parasitoid attraction to cues that guide D. longicaudata to the host, and thus unveil volatile organic compounds that might be used in IPM programmes. Female orientation to chemical cues was tested in a Y‐tube olfactometer, where attraction to C. capitata‐infested oranges, oranges with residues of larval activity, oranges infected with a green mould, and overripe oranges was confirmed. Volatiles from all these types of fruit were collected and used in gas chromatography‐electroantennographic detection (GC‐EAD) and GC‐mass spectrometry (MS) analyses. These studies allowed us to identify six candidate compounds that were present in all treated oranges but not in the control fruit (ripe and uninfested oranges): D‐limonene, acetophenone, linalool, nonanal, decanal, and eugenol. Electroantennography (EAG) showed that acetophenone, nonanal, and decanal triggered dose‐dependent responses, suggesting a relevant role in the process of host finding. Although responses to D‐limonene, linalool, and eugenol were independent of the dose, they could be involved in host location in areas with high probability of host presence. The fact that these six compounds are shared by the four behaviourally attractive sources opens new possibilities for the development of novel tools to improve biocontrol programmes.

Subcellular biochemistry and biology of filamentous entomopathogenic fungi.

Filamentous entomopathogenic fungi (EPF) function as important biotic factors regulating the arthropod population in natural ecosystems and have great potential as biocontrol agents in modern agriculture. In the infection cycle, EPF undergo a plethora of physiological processes, including metabolism (e.g., cuticle hydrolysis and nutrient utilization), development (e.g., dimorphism and conidiation), stress response (e.g., oxidative and osmotic stresses), and immune evasion from the host. In-depth explorations of the mechanisms involved in the lifecycle of EPF offer excellent opportunities to increase their virulence and stability, which increases the efficacy of EPF in biocontrol programs. This review discusses the current state of knowledge relating to the biological roles and regulatory mechanisms of organelles and subcellular structures in the physiology of EPF, as well as some suggestions for future investigation.

Utilization of Metarhizium as an insect biocontrol agent and a plant bioinoculant with special reference to Brazil.

Brazil has a long history of using biological control and has the largest program in sugarcane agriculture to which a biocontrol program has been applied. This achievement is at least partly due to the utilization of the entomopathogenic fungus Metarhizium. This well-known fungal genus exhibits pathogenicity against a broad range of arthropod hosts and has been used globally as a biocontrol agent. This fungus is also a root symbiont, and in this capacity, it is a plant growth promoter. However, this feature (i.e., as a plant symbiont) has yet to be fully explored and implemented in Brazil, although the number of reports demonstrating Metarhizium's utility as a plant bioinoculant is increasing. The Brazilian bioproduct industry targets agricultural pests, and is limited to two Metarhizium species represented by four fungal isolates as active ingredients. Entomopathogenic fungi have also been successful in controlling arthropods of public health concern, as shown in their control of mosquitoes, which are vectors of diseases. The isolation of new indigenous Metarhizium isolates from a variety of substrates such as soil, insects, and plants shows the wide genetic diversity within this fungal genus. In this review, we emphasize the significance of Metarhizium spp. for the biological control of insects in Brazil. We also suggest that the experience and success of biological control with fungi in Brazil is an important resource for developing integrated pest management and sustainable strategies for pest control worldwide. Moreover, the future implementation prospects of species of Metarhizium being used as bioinoculants and possible new advances in the utility of this fungus are discussed.

Reproductive Parameters and Host Specificity of Eurytoma erythrinae (Hymenoptera: Eurytomidae), a Biological Control Agent of the Erythrina Gall Wasp, Quadrastichus erythrinae (Hymenoptera: Eulophidae).

Eurytoma erythrinae Gates & Delvare (Hymenoptera: Eurytomidae) is an important biological control agent of the erythrina gall wasp (EGW), Quadrastichus erythrinae Kim (Hymenoptera: Eulophidae), an invasive species likely originating in eastern Africa that is a threat to Erythrina trees in Hawaii and worldwide. Thousands of Erythrina trees in Hawaii have succumbed to EGW since 2005 and died within a few years of infestation. The endemic wiliwili tree, Erythrina sandwicensis, an important component of Hawaii's dry forests and one of few deciduous native trees, were severely impacted by this wasp. Early during the invasion by EGW it became evident that the endemic species may be driven to extinction, and exploration programs for natural enemies of the EGW started in December 2005. East Africa was selected as the starting point for natural enemy exploration owing to high native Erythina species richness. Several gall formers were found in Tanzania and a putative color variant type of Q. erythrinae was detected in association with three ectoparasitoids. During January 2006, the dominant parasitoid of this gall former was introduced to Hawaii and described as the new species, E. erythrinae. It was found in Ghana and South Africa attacking other gall wasp species on Erythrina. Eurytoma erythrinae was a voracious ectoparasitoid feeding as a predator on 1-5 adjacent EGW immatures to complete its development. Host specificity studies that included seven nontarget gall-forming species showed no evidence of attraction or parasitism by this parasitoid. Mean ± SEM longevity of host-deprived females (40.4 ± 2.2 days) was significantly higher than males (20.5 ± 1.1 days). Host feeding enhanced longevity of ovipositing females (51.3 ± 1.5 days). Female E. erythrinae is synovigenic, with high egg-maturation rate. Peak fecundity (105-239 offspring/female), host feeding biology, short life cycle (18.4 ± 0.1 days), and synchronization with the host were additional desirable attributes of this species. The parasitoid was approved for field release in Hawaii in November 2008. A total of 3998 wasps were distributed on six Hawaiian Islands, with establishment in less than a year. Impacts on high density infestations of EGW were sufficient to prevent tree deaths. Limited rates of parasitism on low-density galled leaves, flowers, and seedpods necessitated the consideration for releasing a second parasitoid, Aprostocetus nitens Prinsloo & Kelly (Hymenoptera: Eulophidae). We report on the reproductive characteristics and host specificity of E. erythinae that could be of importance for classical biocontrol programs in areas with an EGW problem.