Bee Larvae Research Articles

Imidacloprid disrupts larval molting regulation and nutrient energy metabolism, causing developmental delay in honey bee Apis mellifera.

Imidacloprid is a global health threat that severely poisons the economically and ecologically important honeybee pollinator, Apis mellifera. However, its effects on developing bee larvae remain largely unexplored. Our pilot study showed that imidacloprid causes developmental delay in bee larvae, but the underlying toxicological mechanisms remain incompletely understood. In this study, we exposed bee larvae to imidacloprid at environmentally relevant concentrations of 0.7, 1.2, 3.1, and 377 ppb. There was a marked dose-dependent delay in larval development, characterized by reductions in body mass, width, and growth index. However, imidacloprid did not affect on larval survival and food consumption. The primary toxicological effects induced by elevated concentrations of imidacloprid (377 ppb) included inhibition of neural transmission gene expression, induction of oxidative stress, gut structural damage, and apoptosis, inhibition of developmental regulatory hormones and genes, suppression of gene expression levels involved in proteolysis, amino acid transport, protein synthesis, carbohydrate catabolism, oxidative phosphorylation, and glycolysis energy production. In addition, we found that the larvae may use antioxidant defenses and P450 detoxification mechanisms to mitigate the effects of imidacloprid. Ultimately, this study provides the first evidence that environmentally exposed imidacloprid can affect the growth and development of bee larvae by disrupting molting regulation and limiting the metabolism and utilization of dietary nutrients and energy. These findings have broader implications for studies assessing pesticide hazards in other juvenile animals.

Imidacloprid disrupts larval molting regulation and nutrient energy metabolism, causing developmental delay in honey bee Apis mellifera

Imidacloprid is a global health threat that severely poisons the economically and ecologically important honeybee pollinator, Apis mellifera. However, its effects on developing bee larvae remain largely unexplored. Our pilot study showed that imidacloprid causes developmental delay in bee larvae, but the underlying toxicological mechanisms remain incompletely understood. In this study, we exposed bee larvae to imidacloprid at environmentally relevant concentrations of 0.7, 1.2, 3.1, and 377 ppb. There was a marked dose-dependent delay in larval development, characterized by reductions in body mass, width, and growth index. However, imidacloprid did not affect on larval survival and food consumption. The primary toxicological effects induced by elevated concentrations of imidacloprid (377 ppb) included inhibition of neural transmission gene expression, induction of oxidative stress, gut structural damage, and apoptosis, inhibition of developmental regulatory hormones and genes, suppression of gene expression levels involved in proteolysis, amino acid transport, protein synthesis, carbohydrate catabolism, oxidative phosphorylation, and glycolysis energy production. In addition, we found that the larvae may use antioxidant defenses and P450 detoxification mechanisms to mitigate the effects of imidacloprid. Ultimately, this study provides the first evidence that environmentally exposed imidacloprid can affect the growth and development of bee larvae by disrupting molting regulation and limiting the metabolism and utilization of dietary nutrients and energy. These findings have broader implications for studies assessing pesticide hazards in other juvenile animals.

Jelleine, a Family of Peptides Isolated from the Royal Jelly of the Honey Bees (Apis mellifera), as a Promising Prototype for New Medicines: A Narrative Review.

The jelleine family is a group of four peptides (jelleines I-IV) originally isolated from the royal jelly of honey bee (Apis mellifera), but later detected in some honey samples. These oligopeptides are composed of 8-9 amino acid residues, positively charged (+2 to +3 at pH 7.2), including 38-50% of hydrophobic residues and a carboxamide C-terminus. Jelleines, generated by processing of the C-terminal region of major royal jelly proteins 1 (MRJP-1), play an important biological role in royal jelly conservation as well as in protecting bee larvae from potential pathogens. Therefore, these molecules present numerous benefits for human health, including therapeutic purposes as shown in preclinical studies. In this review, we aimed to evaluate the biological effects of jelleines in addition to characterising their toxicities and stabilities. Jelleines I-III have promising antimicrobial activity and low toxicity (LD50 > 1000 mg/Kg). However, jelleine-IV has not shown relevant biological potential. Jelleine-I, but not the other analogues, also has antiparasitic, healing, and pro-coagulant activities in addition to indirectly modulating tumor cell growth and controlling the inflammatory process. Although it is sensitive to hydrolysis by proteases, the addition of halogens increases the chemical stability of these molecules. Thus, these results suggest that jelleines, especially jelleine-I, are a potential target for the development of new, effective and safe therapeutic molecules for clinical use.

Quantitative tyramine analysis method for Apis mellifera larvae infected with Melissococcus plutonius, the causative agent of European foulbrood.

Tyramine, a trace monoamine produced from tyrosine by decarboxylation and found naturally in foods, plants, and animals, is a suspected virulence factor of Melissococcus plutonius that causes European foulbrood in honey bee brood. In the present study, we developed a method for quantitative analysis of tyramine in culture medium and honey bee larvae with a limit of quantitation of 3 ng/mL and a recovery rate of >97% using Liquid Chromatography-Mass Spectrometry/Mass Spectrometry and deuterium-labeled tyramine, demonstrating for the first time that a highly virulent M. plutonius strain actually produces tyramine in infected larvae. This method will be an indispensable tool to elucidate the role of tyramine in European foulbrood pathogenesis in combination with exposure bioassays using artificially reared bee larvae.

Microbiomes, diet flexibility, and the spread of a beetle parasite of honey bees.

Invasive pests may disturb and destructively reformat the local ecosystem. The small hive beetle (SHB), Aethina tumida, originated in Africa and has expanded to America, Australia, Europe, and Asia. A key factor facilitating its fast global expansion is its ability to subsist on diverse food inside and outside honey bee colonies. SHBs feed on various plant fruits and exudates in the environment while searching for bee hives. After sneaking into a bee hive, they switch their diet to honey, pollen, and bee larvae. How SHBs survive on such a broad range of food remains unclear. In this study, we simulated the outside and within hive stages by providing banana and hive resources and quantified the SHB associated microbes adjusted by the diet. We found that SHBs fed on bananas were colonized by microbes coding more carbohydrate-active enzymes and a higher alpha diversity than communities from SHBs feeding on hive products or those collected directly from bee hives. SHBs fed on bananas and those collected from the hive showed high symbiont variance, indicated by the beta diversity. Surprisingly, we found the honey bee core symbiont Snodgrassella alvi in the guts of SHBs collected in bee hives. To determine the role of S. alvi in SHB biology, we inoculated SHBs with a genetically tagged culture of S. alvi, showing that this symbiont is a likely transient of SHBs. In contrast, the fungus Kodamaea ohmeri is the primary commensal of SHBs. Diet-based microbiome shifts are likely to play a key role in the spread and success of SHBs.

Hedonic rating coupled with sensory profiles using CATA for six whole roasted or deep-fried insects among Danish 11-13-year-old children

The consumption of insects is not rooted in traditional Western culture, which creates a barrier to integrate insects into our diets, especially among preadolescents. This study was executed during a Danish science outreach week to educate children on eating insects through educational activities. 11–13-y.o. children were invited to taste six different types of whole roasted or deep-fried insects (grasshoppers, cricket, bee larvae, wax moth larvae, buffalo worms and mealworms), asked to rate their liking (15-point hedonic scale) and describe the sensory properties for each insect (50 sensory attributes using Check-All-That-Apply format). Food neophobia was assessed using the Food Neophobia Test Tool to explore the link between food neophobia and hedonic responses of insects. Results showed significant differences in sensory and hedonic responses (p < 0.0001, respectively) between insect types. In general, insects showed low mean liking. They were distributed in two significantly different groups. Mealworms (6.9), buffalo worms (6.6), and bee larvae (6.4) had higher mean liking compared to crickets (5.2), wax moth larvae (5.1), and grasshoppers (4.7). Furthermore, 19 descriptors discriminated between insects demonstrating that insects vary in sensory characteristics. “Crunchy” was significantly associated with liking, highlighting the importance of this sensory attribute when promoting the consumption of edible insects in preadolescents. Insect liking decreased with increasing levels of food neophobia, which was also affected by age- and gender. The study offers new insights into the acceptance and sensory properties of commercial insects that can be used to promote insect consumption and for future product development targeted at this group.

Detection of Ascosphaera apis, causing chalkbrood disease in the colonies of European honey bee, Apis mellifera in West Bengal, India

The decline of honey bee (Apis mellifera L.) populations is of great concern around the world. Among the several key drivers, dissemination of pests and pathogens is potential one. Chalkbrood is very common fungal disease of honey bee, caused due to Ascosphaera apis. In the present study, survey was conducted regarding the prevalence of diseases in A. mellifera beekeeping in Gangetic plains of West Bengal, India. Results confirmed the occurrence of chalkbrood disease in different apiaries, where dead and mummified bee larvae with cotton like chalky white or greyish-black covering were found as physical symptoms of the disease. From three surveyed apiaries, a total of 16 hives out of 113 hives were found to be infected with the diseases; and 46 frames out of 132 frames from the infected 16 hives were found to be affected by the pathogen. Microscopic examination reflected that nearly 87% of the samples collected from the infected frame were found to be positive for the spore of Ascosphaera. The fungus associated with the disease was isolated on Potato Dextrose Agar medium, pure cultured and its genomic DNA was isolated to perform PCR and based on 18s rDNA sequencing by using specific primer pair of ITS-1 and ITS-4, the fungus was identified as Ascosphaera apis. &#x0D; Keywords: 18s rDNA, Bees, Necrotrophs, Phylogeny

A systemic fungicide might reduce the male genitalia of a stingless bee species by one-third

Bees are essential pollinating insects that significantly contribute to crop production. However, the use of pesticides in modern agriculture has resulted in bees being exposed to a plethora of harmful substances. Larvae of bees are particularly susceptible to exposure, as they can consume contaminated larval food during development. This study opportunely examined the effects of fungicides and insecticides on the size and shape of the genitalia of Scaptotrigona bipunctata (Lepeletier, 1836) males (Hymenoptera: Apidae: Meliponini) during larval development when experimental design was structured to obtain female bees (workers). Therefore, the geometric morphometric analyses were based on male bees that opportunistically emerged. Our findings showed a significant difference in the size of the genitalia of S. bipunctata males exposed to a systemic fungicide during larval development, while the genital shape remained similar. Although the exact impact of these morphological changes on the reproductive success of S. bipunctata males is uncertain, they suggest an adverse effect of pesticides on bees.

Early-Life Sublethal Thiacloprid Exposure to Honey Bee Larvae: Enduring Effects on Adult Bee Cognitive Abilities.

Honey bees have significant ecological and economic value as important pollinators, but they are continuously exposed to various environmental stressors, including insecticides, which can impair their health and cause colony decline. (1) Background: Cognitive abilities are vital for the functional maintenance of honey bees; however, it remains unknown if chronic, low-dose exposure to thiacloprid during the larval stage impairs the cognitive abilities of emerged adult honey bees. (2) Methods: To explore this question, honey bee larvae were fed 0, 0.5, and 1.0 mg/L thiacloprid during their developmental phase. Then, the cognitive (i.e., olfactory learning and memory) abilities of adult honey bees were quantified to assess the delayed impacts of early-stage thiacloprid exposure on adult honey bee cognition. Neural apoptosis and transcriptomic level were also evaluated to explore the neurological mechanisms underlying these effects. (3) Results: Our results revealed that chronic larval exposure to sublethal thiacloprid impaired the learning and memory abilities of adult honey bees by inducing neuronal apoptosis and transcriptomic alterations. (4) Conclusions: We highlighted a previously unknown impairment caused by thiacloprid in honey bees.

New insights into changing honey bee (Apis mellifera) immunity molecules pattern and fatty acid esters, in responses to Ascosphaera apis infection

Monitoring of metabolite changes could provide valuable insights into disturbances caused by an infection and furthermore, could be used to define the status of an organism as healthy or diseased and define what could be defensive elements against the infection. The present investigation conducted a gas chromatography–mass spectrometry (GC/MS) for haemolymph of larval honey bees (Apis mellifera L.) infected with the fungal pathogen Ascosphaera apis in comparison with control haemolymph non-infected insects. Results revealed that the pathogen caused a general disturbance of metabolites detected in the haemolymph of the honey bee. The majority of metabolites identified before and after infection were fatty acid esters. The disease caused an elevation in levels of methyl oleate, methyl palmitate, and methyl stearate, respectively. Further, the disease drove to the disappearance of methyl palmitoleate, and methyl laurate. Conversely, methyl linolelaidate, and ethyl oleate were identified only in infected larvae. A high reduction in diisooctyl phthalate was recorded after the infection. Interestingly, antimicrobial activities were confirmed for haemolymph of infected honey bee larvae. In spite of the presence of some previously known bioactive compounds in healthy larvae there were no antimicrobial activities.

The Juvenile-Hormone-Responsive Factor AmKr-h1 Regulates Caste Differentiation in Honey Bees.

Honey bees are typical model organisms for the study of caste differentiation, and the juvenile hormone (JH) is a crucial link in the regulatory network of caste differentiation in honey bees. To investigate the mechanism of JH-mediated caste differentiation, we analyzed the effect of the JH response gene AmKr-h1 on this process. We observed that AmKr-h1 expression levels were significantly higher in queen larvae than in worker larvae at the 48 h, 84 h, and 120 h larval stages, and were regulated by JH. Inhibiting AmKr-h1 expression in honey bee larvae using RNAi could lead to the development of larvae toward workers. We also analyzed the transcriptome changes in honey bee larvae after AmKr-h1 RNAi and identified 191 differentially expressed genes (DEGs) and 682 differentially expressed alternative splicing events (DEASEs); of these, many were related to honey bee caste differentiation. Our results indicate that AmKr-h1 regulates caste differentiation in honey bees by acting as a JH-responsive gene.

Importance of pollinator diversity for sunflower seed production

The sunflower (Helianthus annuus Linnaeus) is one of the most important food crops in the world. In modern agroecology, this pivotal plant is deeply intertwined with diverse pollination mechanisms. This study elucidates the multifaceted contributions of various pollinator taxa, delving into their respective efficiencies and behaviors vis-à-vis sunflower reproductive success. The honeybee (Apis mellifera Linnaeus, 1758) plays a central role in sunflower pollination, transferring pollen over large distances, thereby bolstering the genetic diversity of sunflower populations. While sunflowers derive immense benefits from honeybee pollination, they reciprocally offer their pollen as vital nutrition for bee larvae. However, this symbiotic relationship is nuanced, with honeybees occasionally exhibiting selectivity based on sunflower cultivar. While honeybees are paramount, the richness of the ecosystem is further accentuated by the presence of other pollinators, each contributing uniquely to the sunflower’s lifecycle. The foraging fidelity of solitary bees, such as the leafcutter bee (Megachile spp.) and mason bee (Osmia spp.), underscores their indispensable role in cross-pollination. Bumblebees (Bombus spp.) introduce a fascinating "buzz pollination" mechanism, leveraging vibrations to optimize pollen release. Simultaneously, wasps provide functions beyond pollination, serving as biocontrol agents by preying on detrimental pests. Surprisingly, mosquitoes appeared to also be contributors to pollination, especially when traditional pollinators are scarce, with their nocturnal tendencies augmenting pollination continuity. Hoverflies render dual benefits: they aid in pollination and their larvae predate on pests. While butterflies are primarily nectar-oriented, they inadvertently partake in pollen transfer. Moths, through their nocturnal activities, complement pollination. Beetles, often overlooked, act as secondary pollinators. Regrettably, contemporary agricultural practices often disrupt this ecological equilibrium. Pervasive pesticide use and habitat fragmentation imperil these pollinators, underscoring the urgent need for pollinator-friendly approaches. As climate conditions fluctuate, understanding these pollinator dynamics becomes increasingly critical. This comprehensive examination advocates for holistic conservation strategies, targeting sustained sunflower yields and broader ecosystem resilience.

No impact of hygienic behavior and viral coinfection on the development of European foulbrood in honey bee (Apis mellifera) colonies during blueberry pollination in Michigan.

European foulbrood (EFB) is a severe disease of honey bee (Apis mellifera) larvae caused by the bacterium Linnaeus [Hymenoptera: Apidae]) Melissococcus plutonius (ex White) Bailey and Collins (Lactobacillales: Enterococcaceae). Many beekeepers in North America report severe EFB following blueberry pollination, but it is not clear what factors during pollination are related to clinical disease. Additionally, the impact that other factors such as viral load and hygienic behavior have on EFB has not been studied. In Spring of 2020 we enrolled 60 commercial honey bee colonies in a prospective cohort study. Colonies were inspected 3 times over the season with hive metrics and samples taken for viral testing. Each colony was tested for hygienic behavior twice and the score was averaged. Viral loads were determined by qPCR for deformed wing virus (DWV) A and B. We found no statistical difference in the EFB prevalence or severity between the 2 yards at any timepoint; 50% (n = 16) of the colonies in the holding yard and 63% (n = 17) in blueberry developed moderate to severe EFB over the study period. When colonies from both yards were pooled, we found no relationship between viral load or hygienic behavior and development of EFB. These results suggest that other factors may be responsible for driving EFB virulence and hygienic behavior is not likely helpful in managing this disease.

Substances in the mandibular glands mediate queen effects on larval development and colony organization in an annual bumble bee.

Social organization is commonly dynamic, with extreme examples in annual social insects, but little is known about the underlying signals and mechanisms. Bumble bee larvae with close contact to a queen do not differentiate into gynes, pupate at an earlier age, and are commonly smaller than siblings that do not contact a queen. We combined detailed observations, proteomics, microRNA transcriptomics, and gland removal surgery to study the regulation of brood development and division of labor in the annual social bumble bee Bombus terrestris. We found that regurgitates fed to larvae by queens and workers differ in their protein and microRNA composition. The proteome of the regurgitate overlaps significantly with that of the mandibular (MG) and hypopharyngeal glands (HPG), suggesting that these exocrine glands are sources of regurgitate proteins. The proteome of the MG and HPG, but not the salivary glands, differs between queens and workers, with caste-specificity preserved for the MG and regurgitate proteomes. Queens subjected to surgical removal of the MG showed normal behavior, brood care, and weight gain, but failed to shorten larval development. These findings suggest that substances in the queen MG are fed to larvae and influence their developmental program. We suggest that when workers emerge and contribute to larval feeding, they dilute the effects of the queen substances, until she can no longer manipulate the development of all larvae. Longer developmental duration may allow female larvae to differentiate into gynes rather than to workers, mediating the colony transition from the ergonomic to the reproductive phase.

Can larval Osmia bees (Megachilidae) tolerate conifer resin volatiles?

Some bees of the Megachilidae, particularly anthidiines, collect plant resins to partition, plug and sometimes line their nest cells. Adult females possess putative morphological, behavioral and perhaps exocrine adaptations for resin harvest, transport and handling. Their eggs and larvae later experience volatiles from the degassing resin, but are they specially adapted to tolerate or detoxify the monoterpenes and other volatiles? To answer this question, eggs and larvae of a megachilid mason bee that builds with mud, Osmia lignaria, were chronically exposed to volatiles emanating from fresh pine and spruce resins placed in their closed brood cells. Although their eggs and larvae do not normally encounter volatiles emanating from pine or spruce resin, they survived without ill effect. Therefore, no adaptive hypothesis needs to be invoked to explain resin volatile tolerance by immatures of resin-using anthidiine and Megachile bees. I briefly discuss potential implications for some other brood cell occupants.

Effects of the insecticide imidacloprid on the post-embryonic development of the honey bee Apis mellifera (Hymenoptera: Apidae)

The widespread use of pesticides in agriculture has been linked to declines in bee populations worldwide. Imidacloprid is a widely used systemic insecticide that can be found in the pollen and nectar of plants and has the potential to negatively impact the development of bee larvae. We investigated the effects of oral exposure to a realistic field concentration (20.5 ng g−1) of imidacloprid on the midgut and fat body of Apis mellifera worker larvae. Our results showed that larvae exposed to imidacloprid exhibited changes in the midgut epithelium, including disorganization of the brush border, nuclear chromatin condensation, cytoplasm vacuolization, and release of cell fragments indication cell death. Additionally, histochemical analysis revealed that the midgut brush border glycocalyx was disorganized in exposed larvae. The fat body cells of imidacloprid-exposed larvae had a decrease in the size of lipid droplets from 50 to 8 μm and increase of 100 % of protein content, suggesting possible responses to the stress caused by the insecticide. However, the expression of de cdc20 gene, which plays a role in cell proliferation, was not affected in the midgut and fat body of treated larvae. These results suggest that imidacloprid negatively affects non-target organs during the larval development of A. mellifera potentially impacting this important pollinator species.

Age matters: variations in parasitoid diversity along a successional gradient in a dry semi-deciduous tropical forest.

Parasitoids are an important group of insects because their species number is among the highest. Multiple studies have addressed the relationships between forest successional age and insect diversity by focusing on herbivorous organisms, but changes in diversity of parasitoids are still poorly known. This work analyses the diversity of parasitoids in tropical forests representing three successional stages. A total of 30 traps were placed, ten in each forest successional stages. We estimated true diversity of Ichneumonidae species and guilds and explored the relationship between their diversity and the abundance of plant species using an Indicator Species Analysis; the relationship between parasitoid species and plant richness and abundance was tested using a Redundancy Analysis. A total of 1522 individuals and 168 morpho-species were captured in four months. Species richness showed no differences; however, parasitoid abundance was higher in young forest, while intermediate forest had the highest true diversity values (1D) with 71.6 effective species. According to insect guilds, richness, abundance, and diversity were similar in the three vegetation successional stages. This finding may be explained based on the intermediate disturbance hypothesis, which postulates that moderate disturbance levels favor the highest diversity. In conclusion, successional age matters, i.e., diversity is the highest in intermediate stages, while the old forests harbors guilds unique to that successional stage, such as parasitoids of melitophagous larvae of bees. Other successional stages were characterized by a single species of parasitoid, belonging to the genera Eiphosoma and Anomalon, which may indicate altered and preserved forests, respectively.

Honey Bee Larval Hemolymph as a Source of Key Nutrients and Proteins Offers a Promising Medium for Varroa destructor Artificial Rearing.

Varroa destructor, a major ectoparasite of the Western honey bee Apis mellifera, is a widespread pest that damages colonies in the Northern Hemisphere. Throughout their lifecycle, V. destructor females feed on almost every developmental stage of their host, from the last larval instar to the adult. The parasite is thought to feed on hemolymph and fat body, although its exact diet and nutritional requirements are poorly known. Using artificial Parafilm™ dummies, we explored the nutrition of V. destructor females and assessed their survival when fed on hemolymph from bee larvae, pupae, or adults. We compared the results with mites fed on synthetic solutions or filtered larval hemolymph. The results showed that the parasites could survive for several days or weeks on different diets. Bee larval hemolymph yielded the highest survival rates, and filtered larval plasma was sufficient to maintain the mites for 14 days or more. This cell-free solution therefore theoretically contains all the necessary nutrients for mite survival. Because some bee proteins are known to be hijacked without being digested by the parasite, we decided to run a proteomic analysis of larval honey bee plasma to highlight the most common proteins in our samples. A list of 54 proteins was compiled, including several energy metabolism proteins such as Vitellogenin, Hexamerin, or Transferrins. These molecules represent key nutrient candidates that could be crucial for V. destructor survival.

Glyphosate affects larval gut microbiota and metamorphosis of honey bees with differences between rearing procedures

The honey bee Apis mellifera is a sentinel species of the pollinator community which is exposed to a wide variety of pesticides. In the last half-century, the pesticide most applied worldwide has been the herbicide glyphosate (GLY) used for weed control and with microbiocide effects. After its application in crops, the GLY residues have been detected in flowers visited by honey bees as well as in the stored food of their hives. Therefore, the honey bee brood can ingest the herbicide during larval development. Recent studies proved that GLY has detrimental effects on adult honey bees and other insects associated with the disturbance of their gut microbiota. GLY induces changes in the growth, metabolism and survival of honey bees and stingless bees reared in vitro. However, the effect of GLY on larval microbiota is unknown so far and there are few studies with an in-hive exposure to GLY. For these reasons, this study aims to determine whether GLY induces dysbiosis in honey bee larvae and affects their metamorphosis during the exposure period (pre-defecation) and the post-exposure period. Furthermore, we assessed this herbicide in vitro and in the hive to compare its effects on different rearing procedures. Finally, we tested the pigment BLUE1 as an indirect exposure marker to detect and estimate the in-hive intake concentration of GLY. Our results indicate that the intake of field-relevant concentrations of GLY induced a slowdown in growth with dysbiosis in the larval gut microbiota followed by late effects on their metamorphosis such as teratogenesis and mortality of newly emerged bees. Nevertheless, brood from the same colonies expressed different signs of toxicity depending on the rearing procedure and in a dose-dependent manner.

Effects of fungicides propiconazole and Trichoderma spp. on the mortality and the physiological status of larvae and adult worker honey bees (Apis mellifera L.)

Exposure to pesticides may affect the survival and normal physiological functions of the honey bee (Apis mellifera L.). The fungicide propiconazole (PRO) and the biofungicide Trichoderma are widely used in agroecosystems, and their effects on the physiology of the honey bee have been largely neglected in risk assessment procedures. We investigated the mortality and variations in physiological biomarkers of honey bee larvae and newly emerged workers exposed to PRO and Trichoderma. Exceeding the field-recommended concentrations of Trichoderma decreased the survival of newly emerged bee workers significantly through oral exposure. PRO, however, showed no significant lethal effects. Contact or oral exposure to field-realistic concentrations of PRO and Trichoderma profoundly altered the activities of detoxification, antioxidative, and digestive enzymes along with energy reserve concentrations of honey bee larvae and adult workers. Integrated biological index (IBRv2) values suggested potential risks of PRO and Trichoderma to honey bees by impairing the physiological status of larvae and newly emerged workers.