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Pathogen infection and conspecific density may considerably affect key life‐history traits of organisms. For naturally aggregating species, even low concentrations of pathogens or high larval densities may have detrimental effects. However, the detailed influence of these factors, particularly their interaction effect, is often overlooked in ecological and life‐history studies. To investigate the effects of conspecific density and pathogen infection on life‐history traits, we explored the influence of larval density (1 and 5 larvae cm−2, i.e., low and high density, respectively) on phenoloxidase (PO) activity, body mass, and development time of Hermetia illucens L. (Diptera: Stratiomyidae) larvae in the presence of two strains of the entomopathogenic fungus (EPF) Beauveria bassiana (Bals.‐Criv.) Vuill. We observed higher PO activity in EPF‐treated larvae than in the untreated control and a pronounced difference in PO activity between the two EPF‐strain treatments. Larvae reared at high density and treated with EPF showed higher PO activity than untreated larvae at low density. The EPF‐treated larvae and larvae reared at high density had longer larval periods than untreated larvae and larvae reared at low density, respectively. Larvae reared at high density also achieved reduced prepupal and pupal masses compared to conspecifics at low density. Interestingly, untreated larvae only achieved higher prepupal and pupal masses at low density, whereas at high density, the pattern was reversed (treated individuals had higher pupal masses). Overall, our results demonstrate that high density and fungal pathogens both induce a higher immune response compared to low density and pathogen‐free environments, but this comes with a cost of a longer larval period and reduced body mass. [ABSTRACT FROM AUTHOR]

Simple Summary: Several lab studies have shown that pesticides are causing sublethal effects on bees; however, little is known about their impact in actual field conditions. Continuous surveillance of the bees in the field will help monitor their pesticide exposure and better understand their impact on bee health. This study investigated the potential use of a new optical sensor, coupled with machine learning, to automatically identify flying insects, as an approach to monitoring the sublethal effects of pesticides on insects. To do so, bumblebees, Bombus terrestris, were exposed to field-realistic doses of a neonicotinoid pesticide, and their flight activity was recorded by the sensor. Through the data acquired, the investigation focused on whether the machine learning algorithm could distinguish the flight events that were created by the healthy bumblebees and the bumblebees that had been exposed to sublethal doses of the pesticide. The results showed that the algorithm could differentiate flight events, indicating the promise of the sensor as a continuous monitoring tool for bee health in the field. Among insects, bees are important pollinators, providing many vital ecosystem services. The recent pollinator decline is threatening both their diversity and abundance. One of the main drivers of this decline is the extensive use of pesticides. Neonicotinoids, one of the most popular groups of pesticides, can be toxic to bees. In fact, numerous studies have found that neonicotinoids can cause sublethal effects, which can impair the biology, physiology, and colony survival of the bees. Yet, there are still knowledge gaps, and more research is needed to better understand the interaction between neonicotinoids and bees, especially in the field. A new optical sensor, which can automatically identify flying insects using machine learning, has been created to continuously monitor insect activity in the field. This study investigated the potential use of this sensor as a tool for monitoring the sublethal effects of pesticides on bumblebees. Bombus terrestris workers were orally exposed to field-realistic doses of imidacloprid. Two types of exposures were tested: acute and chronic. The flight activity of pesticide-exposed and non-exposed bumblebees was recorded, and the events of the insect flights recorded by the sensor were used in two ways: to extract the values of the wingbeat frequency and to train machine learning models. The results showed that the trained model was able to recognize differences between the events created by pesticide-exposed bumblebees and the control bumblebees. This study demonstrates the possibility of the optical sensor for use as a tool to monitor bees that have been exposed to sublethal doses of pesticides. The optical sensor can provide data that could be helpful in managing and, ideally, mitigating the decline of pollinators from one of their most major threats, pesticides. [ABSTRACT FROM AUTHOR]

Insect production offers a sustainable source of nutrients for livestock. This comes with a risk for transmission of pathogens from the insects into the livestock sector, including viruses causing serious diseases, such as African swine fever virus (ASFV), classical swine fever virus and foot-and-mouth disease virus. ASFV is known to survive for a long time within animal meat and byproducts. Therefore, we conducted experimental exposure studies of insects to ASFV using larvae of two key insect species produced for food and feed, the mealworm; Tenebrio molitor, and the black soldier fly, Hermetia illucens. The larvae were exposed to ASFV POL/2015/Podlaskie, via oral uptake of serum or spleen material from ASFV-infected pigs. Using qPCR, the amounts of viral DNA present immediately after exposure varied from ~104.7 to 107.2 genome copies per insect. ASFV DNA was detectable in the larvae of H. illucens for up to 3 days post exposure and in T. molitor larvae for up to 9 days post exposure. To assess the presence of infectious virus within the larvae and with this, the risk of virus transmission via oral consumption, pigs were fed cakes containing larvae exposed to ASFV. Pigs that consumed 50 T. molitor or 50 H. illucens virus-exposed larvae did not become infected with ASFV. Thus, it appears, that in our experimental setting, the risk of ASFV transmission via consumption of unprocessed insect larvae, used as feed, is low. [ABSTRACT FROM AUTHOR]

Simple Summary: Insects have been widely studied as a potential sustainable source of proteins to meet a rising global demand. Among them, the yellow mealworm (Tenebrio molitor, L.) is showing promise for its mass-rearing potential and its authorization by the European Union (EU) as a novel food. As in conventional animal husbandry practices, probiotics could provide nutritional and immunological benefits as part of the insect's diet. This study evaluated the dietary supplementation of three types of probiotics on the development and disease resistance of yellow mealworm larvae. The results showed that the addition of probiotics can play a role in insect farming to improve the nutritional value of sub-optimal diets and protect the insects against entomopathogens. However, this study emphasizes the contrasting effects of the different probiotic strains tested and the need for more research on the topic. In recent years, the yellow mealworm (Tenebrio molitor L.) has demonstrated its potential as a mass-produced edible insect for food and feed. However, challenges brought on by pathogens in intensive production systems are unavoidable and require the development of new solutions. One potential solution is the supplementation of probiotics in the insect's diet to obtain the double benefits of improved growth and enhanced immune response. The aim of this study was to evaluate the effects of diet-based probiotic supplementation on T. molitor larval survival, growth, and resistance against a fungal pathogen. Three probiotic strains, namely Pediococcus pentosacceus KVL-B19-01 isolated from T. molitor and two commercialized strains for traditional livestock, Enterococcus faecium 669 and Bacillus subtilis 597, were tested. Additionally, when larvae were 9 weeks old, a pathogen challenge experiment was conducted with the fungus Metarhizium brunneum. Results showed that both P. pentosaceus and E. faecium improved larval growth and larval survival following fungal exposure compared to the non-supplemented control diet. Since B. subtilis did not improve larval performance in terms of either development or protection against M. brunneum, this study suggests the need for further research and evaluation of probiotic strains and their modes of action when considered as a supplement in T. molitor's diet. [ABSTRACT FROM AUTHOR]

Agricultural and apicultural practices expose honeybees to a range of pesticides that have the potential to negatively affect their physiology, neurobiology, and behavior. Accumulating evidence suggests that these effects extend to the honeybee gut microbiome, which serves important functions for honeybee health. Here we test the potential effects of the pesticides thiacloprid, acetamiprid, and oxalic acid on the gut microbiota of honeybees, first in direct in vitro inhibition assays and secondly in an in vivo caged bee experiment to test if exposure leads to gut microbiota community changes. We found that thiacloprid did not inhibit the honeybee core gut bacteria in vitro , nor did it affect overall community composition or richness in vivo. Acetamiprid did also not inhibit bacterial growth in vitro , but it did affect community structure within bees. The eight bacterial genera tested showed variable levels of susceptibility to oxalic acid in vitro. In vivo , treatment with this pesticide reduced amplicon sequence variant (ASV) richness and affected gut microbiome composition, with most marked impact on the common crop bacteria Lactobacillus kunkeei and the genus Bombella. We conducted network analyses which captured known associations between bacterial members and illustrated the sensitivity of the microbiome to environmental stressors. Our findings point to risks of honeybee exposure to oxalic acid, which has been deemed safe for use in treatment against Varroa mites in honeybee colonies, and we advocate for more extensive assessment of the long-term effects that it may have on honeybee health. [ABSTRACT FROM AUTHOR]

Black soldier fly [BSF; Hermetia illucens L. (Diptera: Stratiomyidae)] larvae are very effective in transforming low‐grade food waste into valuable high‐end proteins and fat, in intensive production facilities. The production output of this species is growing quickly, but upscaling brings risks to the health status of the reared insects. Until now, not a single major case of disease outbreak caused by a pathogen in a BSF production unit has been reported. This contrasts with data on other species of mass‐produced insects, which have experienced various disease outbreaks, indicating that BSFs are comparatively resistant to insect diseases. Further, there are no records of natural infections caused by entomopathogens in BSF. In this review, the known entomopathogens of Diptera, especially BSF, and their potential risks for causing disease in these insects are summarized. [ABSTRACT FROM AUTHOR]

Entomophthoralean fungi are insect pathogenic fungi and are characterized by their active discharge of infective conidia that infect insects. Our aim was to study the effects of temperature on the discharge and to characterize the variation in the associated temporal pattern of a newly discovered Pandora species with focus on peak location and shape of the discharge. Mycelia were incubated at various temperatures in darkness, and conidial discharge was measured over time. We used a novel modification of a statistical model (pavpop), that simultaneously estimates phase and amplitude effects, into a setting of generalized linear models. This model is used to test hypotheses of peak location and discharge of conidia. The statistical analysis showed that high temperature leads to an early and fast decreasing peak, whereas there were no significant differences in total number of discharged conidia. Using the proposed model we also quantified the biological variation in the timing of the peak location at a fixed temperature. [ABSTRACT FROM AUTHOR]

In temperate regions, insect pathogenic fungi face the challenge of surviving through the winter. Winter is a time when hosts are immobile, low in number or are present in a stage which is not susceptible to infection. Fungi from Entomophthoromycota have so far been known to survive the winter in two ways: either as (1) thick-walled resting spores released into environment from dead hosts, or as (2) structures inside the dead host (e.g., hyphal bodies). Here we report, from the Danish environment, a third way to survive the winter, namely a slow progression and transmission of Entomophthora schizophorae in adult dipteran Pollenia hosts that hibernate in clusters in unheated attics, sheltered areas outdoors (under bark etc.). Fungus-killed sporulating flies were observed outside very early and very late in the season. By sampling adults at the time of their emergence from hibernation in late winter/early spring we documented that the fungus was naturally prevalent and killed flies after a period of incubation. Experimentally we documented that even at the low temperature of 5 °C, the fungus was able to maintain itself in Pollenia cohorts for up to 90 days. From these observations the full winter cycle of this fungus is elucidated. The three types of winter survival are discussed in relation to fungus epidemic development. [ABSTRACT FROM AUTHOR]

The natural occurrence of entomophthoralean fungi pathogenic towards aphids on cereal and potato crops was investigated in the years 2009, 2010, and 2011. Infected aphids were sampled in three bioclimatic zones in Tunisia (Beja, Cap bon, and Kairouan) and fungal species were determined based on morphological characters such as shape, size, and number of nuclei in the primary conidia. Polymerase Chain Reaction (PCR) on the internal transcribed spacer 1 region (ITS1) was used to verify morphological determination. Both methods gave consistent results and we documented for the first time the natural occurrence of two fungal species from the order Entomophthorales (phylum Entomophthoromycota), Pandora neoaphidis and Entomophthora planchoniana. Both fungi were recorded on the aphid species Sitobion avenae and Myzus persicae on barley ears and potato leaves, respectively. Moreover, natural mixed infections by both species (P. neoaphidis and E. planchoniana) were documented on the target aphids. This investigation provides basic information of entomopathogenic fungi infecting economically important aphids in Tunisia. [ABSTRACT FROM AUTHOR]

Ascosphaera subglobosa (Eurotiomycetes: Onygenales) is newly described from the pollen provisions and nesting material of the solitary leaf-cutting bee Megachile rotundata in Canada and the western United States. This new species, related to A. atra and A. duoformis, is distinguished from other Ascosphaera species by its globose to subglobose ascospores, evanescent spore balls and unique nuclear ribosomal DNA sequences (ITS and LSU). [ABSTRACT FROM AUTHOR]

Three DNA regions (ITS 1, LSU rRNA and GPD) of isolates from the insect-pathogenic fungus genus Entomophthora originating from different fly ( Diptera) and aphid ( Hemiptera) host taxa were sequenced. The results documented a large genetic diversity among the fly-pathogenic Entomophthora and only minor differences among aphid-pathogenic Entomophthora. The evolutionary time of divergence of the fly and the aphid host taxa included cannot account for this difference. The host-driven divergence of Entomophthora, therefore, has been much greater in flies than in aphids. Host-range differences or a recent host shift to aphid are possible explanations. [ABSTRACT FROM AUTHOR]

Provides information on a study that explored the association between members of the entomophthoralean genera Eryniopsis and Entomophaga with polymerase chain reaction-restriction fragment length polymorphism using entomophthoralean-specific primers. Methodology of the study; Results and discussion on the study.

We compared nutrient compositions of honey bee (Apis mellifera) drones of different developmental stages from two different populations—the Italian honey bee reared in Korea and Buckfast bees from Denmark. Analyses included amino acid, fatty acid, and mineral content as well as evaluations of antioxidant properties and haemolysis activities. The compositions of total amino acids, and thus protein content of the insects, increased with development. A similar trend was observed for minerals presumably due to the consumption of food in the adult stage. In contrast, total fatty acid amounts decreased with development. Altogether, seventeen amino acids, including all the essential ones, except tryptophan, were determined. Saturated fatty acids dominated over monounsaturated fatty acids in the pupae, but the reverse held true for the adults. Drones were found to be rich in minerals and the particularly high iron as well as K/Na ratio was indicative of the nutritional value of these insects. Among the three developmental stages, adult Buckfast drones exhibited the highest antioxidant activity. Bearing in mind the overall high nutritional value, i.e., high amino acids, minerals and less fatty acids, late pupae and adult drones can be useful for human consumption while the larvae or early pupal stage can be recommended as feed. However, owing to their relatively high haemolysis activity, we advocate processing prior to the consumption of these insects. [ABSTRACT FROM AUTHOR]

Pandora neoaphidis and Entomophthora planchoniana (phylum Entomophthoromycota) are important fungal pathogens on cereal aphids, Sitobion avenae and Rhopalosiphum padi. Here, we evaluated and compared for the first time the virulence of these two fungi, both produced in S. avenae cadavers, against the two aphid species subjected to the same exposure. Two laboratory bioassays were carried out using a method imitating entomophthoralean transmission in the field. Healthy colonies of the two aphid species were exposed to the same conidial shower of P. neoaphidis or E. planchoniana, in both cases from a cadaver of S. avenae. The experiments were performed under LD 18:6 h at 21 °C and a successful transmission was monitored for a period of nine days after initial exposure. Susceptibility of both S. avenae and R. padi to fungal infection showed a sigmoid trend. The fitted nonlinear model showed that the conspecific host, S. avenae, was more susceptible to E. planchoniana infection than the heterospecific host R. padi, was. In the case of P. neoaphidis, LT50 for S. avenae was 5.0 days compared to 5.9 days for R. padi. For E. planchoniana, the LT50 for S. avenae was 4.9 days, while the measured infection level in R. padi was always below 50 percent. Our results suggest that transmission from conspecific aphid host to heterospecific aphid host can occur in the field, but with expected highest transmission success to the conspecific host. [ABSTRACT FROM AUTHOR]

Opportunistic bacteria are often ubiquitous and do not trigger disease in insects unless the conditions are specifically favorable for bacterial development in a suitable host. In this paper, we isolated and identified a bacterium, Pseudomonas aeruginosa, from the larvae of the giant mealworm Zophobas morio and we studied the possible entry routes by challenging larvae with per os injection and subdermal injection. We also evaluated the effect of exposing groups of larvae to P. aeruginosa inoculated in their feed and the effect of exposing wounded larvae to P. aeruginosa. We concluded that the mortality rate of Z. morio larvae is higher when P. aeruginosa gets in direct contact with the hemolymph via intracoelomic injection compared to a situation where the bacterium is force-fed. Larvae with an open wound exposed to P. aeruginosa presented higher mortality rate compared to larvae with a wound that was not exposed to the bacterium. We documented too, that cannibalism and scavenging were more prevalent among larvae in a group, when P. aeruginosa is present compared to when it is absent. We discuss hereby different aspects related with the pathogen's entry routes to insects the complexity of pathogen´s transmission in high population densities and different ways to prevent and/or control P. aeruginosa in mass rearing systems. [ABSTRACT FROM AUTHOR]