Biocontrol Programs Research Articles

Invertebrate and fungal associations of the pastoral weed, Madagascar ragwort, Senecio madagascariensis Poir, in Northland, New Zealand, and implications for biocontrol

ABSTRACT Senecio madagascariensis Poir is now found in New Zealand colonising pastoral habitats north of a latitude 35.5°S. This prolific, continuous flowering, toxic perennial weed may not have reached its potential geographic range in New Zealand and is poorly controlled by herbicides. Prior to embarking on a biocontrol programme for this weed, we present a survey of invertebrates and diseases which found relatively few natural enemies and no specialised herbivores or plant pathogens in its current range in New Zealand. The most significant herbivorous insect pests were Hemipteran sap-suckers from 8 families. Percentage infestation of seedheads by the multivoltine tephritid fly, Sphenella rufriceps (Macquart) ranged from 16.7% to 49.8%. Fewer stems of S. madagascariensis were found in plants infected by the fungus, Puccinia lagenophorae Cooke. A search in Western Province (RSA) and in Madagascar for biocontrol control agents host specific to S. madagascariensis is planned.

From ecological menace to roadside attraction: 28 years of evidence support successful biocontrol of purple loosestrife

AbstractIntroduction and spread of non‐native plants provide ecologists and evolutionary biologists with abundant scientific opportunities. However, land managers charged with preventing ecological impacts face financial and logistical challenges to reduce threats by introduced species. The available toolbox (chemical, mechanical, or biological) is also rather limited. Failure to permanently suppress introduced species by mechanical and chemical treatments may result in biocontrol programs using host‐specific insect herbivores. Regardless of the chosen method, long‐term assessment of management outcomes on both the target species and associated biota should be an essential component of management programs. However, data to assess whether management results in desirable outcomes beyond short‐term reductions of the target plant are limited. Here, we use implementation of a biocontrol program targeting a widespread wetland invader, Lythrum salicaria (purple loosestrife), in North America to track outcomes on the target plant over more than two decades in New York State. After extensive testing, two leaf‐feeding beetles (Galerucella calmariensis and Galerucella pusilla; hereafter “Galerucella”), a root‐feeding weevil (Hylobius transversovittatus) and a flower‐feeding weevil (Nanophyes marmoratus), were approved for field releases. We used a standardized monitoring protocol to record insect abundance and L. salicaria stem densities and heights in 1‐m2 permanent quadrats at 33 different wetlands and followed sites for up to 28 years. As part of this long‐term monitoring, in 20 of these wetlands, we established a factorial experiment releasing either no insects (control), only root feeders, only leaf beetles, or root and leaf feeders. We documented reduced L. salicaria occupancy and stem densities following insect releases over time, irrespective of site‐specific differences in starting plant communities or L. salicaria abundance. We could not complete our factorial experiment because dispersal of leaf beetles to root‐feeder‐only and control sites within 5 years invalidated our experimental controls. Our data show that it took time for significant changes to occur, and short‐term studies may provide misleading results, as L. salicaria stem densities initially increased before significantly decreasing. Several decades after insect releases, prerelease predictions of significant purple loosestrife declines have been confirmed.

Preference behavior of Phytoseiulus persimilis and Neoseiulus californicus affected by heterospecific predators

The initial phase in optimizing the efficacy of multiple predators in biocontrol programs involves elucidating their reactions in the presence of heterospecific competitors. Predators sharing the same trophic level may interfere with one another through competition for food resources and engage in internal predation dynamics. This study explored the compatibility of two predatory mite species, Phytoseiulus persimilis, and Neoseiulus californicus, as biocontrol agents targeting Tetranychus urticae. Plants with prey and those with prey plus heterospecifics were used to assess the preference of adult females of the predatory mite competitors. Our results indicated no significant preference by either predatory mite species for plants with spider mites alone or alongside heterospecifics. Additionally, the fecundity rates of both species remained unaffected by the presence of heterospecifics. Notably, N. californicus strongly preferred aggregating eggs on the side with spider mite webs, contrasting with the more random distribution of P. persimilis eggs, although the distribution of eggs on leaf discs was altered in the presence of higher densities of heterospecifics. The total number of eggs of each predator species remained unaffected by the presence of heterospecifics. While our findings suggest compatibility between P. persimilis and N. californicus, which could enhance their combined use in pest management, further field studies are necessary to validate this potential under diverse environmental conditions and pest densities.

Revealing the larval anatomy of the hoverfly Sphaerophoria rueppellii (Wiedemann, 1820) (Diptera, Syrphidae) using micro-computed tomography

Micro-computed tomography (micro-CT) is an X-ray-based technique that allows visualisation of the internal anatomy of insects in situ and does not require dissections. Traditionally, the study of insect anatomy has been mainly based on dissection techniques and microtome sections. However, micro-CT is becoming an increasingly widespread study technique. We report the use of micro-CT scans to study, in detail, the external and internal structures and organs of the third instar larva of the dipteran Syrphidae Sphaerophoria rueppellii, a Palaearctic species included in the list of effective agents used in European biocontrol programmes against aphids. Detailed images and videos provided evidence of the external morphological characteristics and an overview of the internal anatomy (musculature, digestive tube, salivary glands, Malpighian tubules, fat body, circulatory system, nervous system, cephalopharyngeal complex, tentorium, and mouth parts). The study evidenced the following: an external lateroventral complex of muscles that form muscular rings that allow the compression of the body segments in a similar functional way as occurs in annelid worms; the existence of mandibles with prosthecae; the nervous system present a conspicuous distinguishable suboesophageal ganglion; the so-called antennomaxillary organs are the antennae; the maxillae are separate structures, with an articulated maxillary palpus; S. rueppellii has only one pair of Malpighian tubules instead of the two pairs that were thought to be the general rule for Syrphid larvae; and the evidence of an imaginal disc of genitalia in the posterior part of the body, below the rectum, which in male larvae has a clear correspondence with the morphology of the external male genitalia, which allows to differentiate the sexes in the larval stage. This study constitutes a true anatomical atlas of the third instar larvae of S. rueppellii. High-quality rendered images and additional supplementary videos together with a 3D model, suitable for use with mobile devices, are useful tools for future research and teaching aids.

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.