Honey Bee Apis Mellifera Research Articles

Detrimental effects of amitraz exposure in honey bees (Apis mellifera) infected with Nosema ceranae.

In recent years, there has been growing concern on the potential weakening of honey bees and their increased susceptibility to pathogens due to chronic exposure to xenobiotics. The present work aimed to study the effects on bees undergoing an infection by Nosema ceranae and being exposed to a frequently used in-hive acaricide, amitraz. To achieve this, newly emerged bees were individually infected with N. ceranae spores and/or received a sublethal concentration of amitraz in their diets under laboratory conditions. Mortality, food intake, total volume excrement, body appearance, and parasite development were registered. Bees exposed to both stressors jointly had higher mortality rates compared to bees exposed separately, with no difference in the parasite development. An increase in sugar syrup consumption was observed for all treated bees while infected bees fed with amitraz also showed a diminishment in pollen intake. These results coupled with an increase in the total number of excretion events, alterations in behavior and body surface on individuals that received amitraz could evidence the detrimental action of this molecule. To corroborate these findings under semi-field conditions, worker bees were artificially infected, marked, and released into colonies. Then, they were exposed to a commercial amitraz-based product by contact. The recovered bees showed no differences in the parasite development due to amitraz exposure. This study provides evidence to which extent a honey bee infected with N. ceranae could potentially be weakened by chronic exposure to amitraz treatment.

First large-scale study reveals important losses of managed honey bee and stingless bee colonies in Latin America

Over the last quarter century, increasing honey bee colony losses motivated standardized large-scale surveys of managed honey bees (Apis mellifera), particularly in Europe and the United States. Here we present the first large-scale standardized survey of colony losses of managed honey bees and stingless bees across Latin America. Overall, 1736 beekeepers and 165 meliponiculturists participated in the 2-year survey (2016–2017 and 2017–2018). On average, 30.4% of honey bee colonies and 39.6% of stingless bee colonies were lost per year across the region. Summer losses were higher than winter losses in stingless bees (30.9% and 22.2%, respectively) but not in honey bees (18.8% and 20.6%, respectively). Colony loss increased with operation size during the summer in both honey bees and stingless bees and decreased with operation size during the winter in stingless bees. Furthermore, losses differed significantly between countries and across years for both beekeepers and meliponiculturists. Overall, winter losses of honey bee colonies in Latin America (20.6%) position this region between Europe (12.5%) and the United States (40.4%). These results highlight the magnitude of bee colony losses occurring in the region and suggest difficulties in maintaining overall colony health and economic survival for beekeepers and meliponiculturists.

Parasite and virus dynamics in the honeybee Apis mellifera unicolor on a tropical island recently invaded by Varroa destructor.

In La Réunion, the established honeybee subspecies Apis mellifera unicolor, an endemic subspecies of African lineage, is facing considerable challenges. Since the introduction of the Varroa destructor mite in 2017 high colony losses have been recorded. We investigated the dynamics of V. destructor and two viruses, the Deformed Wing Virus (DWV), known to be transmitted by the mite, and the Chronic Bee Paralysis Virus (CBPV), in A. m. unicolor. Colonies from two apiaries located at 300 and 900 m a.s.l were monitored twice for one year without any acaricide treatment. The brood area, V. destructor infestation rates, DWV and CBPV prevalence and load were recorded monthly. A. m. unicolor maintained brood rearing throughout the year. Varroa destructor infestation resulted in high colony mortality (up to 85 %) and high phoretic mite rates (up to 52 mites per hundred bees). The establishment of DWV in colonies occurred after that of V. destructor and the mite infestation rate had a significant effect on the virus prevalence and load. CBPV appeared only transiently throughout the surveys. The data showed that, in tropical colonies with permanent brood rearing, V. destructor and DWV can reach high levels, but are still subject to seasonal variations that appear to be influenced by environmental conditions. This suggests that beekeeping practices could be adapted by favouring sites and periods for transhumance or acaricide treatment.

Eggs sampling as an effective tool for identifying the incidence of viruses in honey bees involved in artificial queen rearing

The Carniolan honey bee (Apis mellifera carnica) plays an essential role in crop pollination, environment diversity, and the production of honey bee products. However, the health of individual honey bees and their colonies is under pressure due to multiple stressors, including viruses as a significant threat to bees. Monitoring various virus infections could be a crucial selection tool during queen rearing. In the present study, samples from all developmental stages (eggs, larvae, pupae, and queens) were screened for the incidence of seven viruses during queen rearing in Slovenia. The screening of a total of 108 samples from five queen breeders was performed by the RT-qPCR assays. The results showed that the highest incidence was observed for black queen cell virus (BQCV), Lake Sinai virus 3 (LSV3), deformed wing virus B (DWV-B), and sacbrood virus (SBV). The highest viral load was detected in queens (6.07 log10 copies/queen) and larvae (5.50 log10 copies/larva) for BQCV, followed by SBV in larvae (5.47 log10 copies/larva). When comparing all the honey bee developmental stages, the eggs exhibited general screening for virus incidence and load in queen mother colonies. The results suggest that analyzing eggs is a good indicator of resilience to virus infection during queen development.

Influence of honey bee (Apis mellifera) breeding on wing venation in Serbia and neighbouring countries.

In order to improve the productivity of honey bees (Apis mellifera), some of their traits are selected by breeding. On one hand, breeding is mainly based on the natural geographical variation of this species; on the other hand, mass production and distribution of artificially selected queens can significantly affect the natural geographic variation of honey bees. In this study, we have compared honey bee wings originating from breeding and non-breeding populations in Serbia. In the comparison, we have also used data from a large area of south-eastern Europe. The wings were measured using the 19 landmarks indicated on the wing images. The coordinates were analysed using the methodology of geometric morphometrics. We found that honey bees obtained from honey bee queen breeder differed in wing venation from surrounding populations, which are under natural selection. Therefore, we argue against including populations under artificial selection in the analysis of the natural geographical variation of honey bees. In our analysis of non-breeding samples, we found that in south-eastern Europe there is continuous variation in wing venation and no clear boundaries between A. m. carnica, A. m. cecropia, and A. m. macedonica.

Bifidobacterium apis sp. nov., isolated from the gut of honeybee (Apis mellifera).

A novel bifidobacterium (designated F753-1T) was isolated from the gut of honeybee (Apis mellifera). Strain F753-1T was characterized using a polyphasic taxonomic approach. Strain F753-1T was phylogenetically related to the type strains of Bifidobacterium mizhiensis, Bifidobacterium asteroides, Bifidobacterium choladohabitans, Bifidobacterium mellis, Bifidobacterium apousia and Bifidobacterium polysaccharolyticum, having 98.4-99.8 % 16S rRNA gene sequence similarities. The phylogenomic tree indicated that strain F753-1T was most closely related to the type strains of B. mellis and B. choladohabitans. Strain F753-1T had the highest average nucleotide identity (94.1-94.5 %) and digital DNA-DNA hybridization (56.3 %) values with B. mellis Bin7NT. Acid production from amygdalin, d-fructose, gentiobiose, d-mannose, maltose, sucrose and d-xylose, activity of α-galactosidase, pyruvate utilization and hydrolysis of hippurate could differentiate strain F753-1T from B. mellis CCUG 66113T and B. choladohabitans JCM 34586T. Based upon the data obtained in the present study, a novel species, Bifidobacterium apis sp. nov., is proposed, and the type strain is F753-1T (=CCTCC AB 2023227T=JCM 36562T=LMG 33388T).

Exploring the effects of the acaricide cyflumetofen on the vital organs of the honey bee Apis mellifera (Hymenoptera: Apidae) workers

Bees are important for maintaining ecosystems, pollinating crops and producing marketable products. In recent years, a decline in bee populations has been reported, with multifactorial causes, including the intensification of pesticide use in agriculture. Among pesticides, cyflumetofen is an insecticide and acaricide used in apple, coffee and citrus crops, whose main pollinator is the honey bee Apis mellifera. Therefore, this bee is a potential target of cyflumetofen during foraging. This study evaluated the histopathological and cytological damage in the midgut, hypopharyngeal glands and fat body of A. mellifera workers exposed to LC50 of cyflumetofen. The midgut epithelium of exposed bees presented cytoplasmic vacuolization, release of vesicles and cell fragments, which indicate autophagy, increased production of digestive enzymes and cell death, respectively. The cytological analysis of the midgut revealed the dilation of the basal labyrinth and the presence of spherocrystals in the digestive cells. The hypopharyngeal glands produced greater amounts of secretion in treated bees, whereas no changes were observed in the fat body. The results indicate that acute exposure to cyflumetofen negatively affect A. mellifera, causing damage to the midgut and changes in the hypopharyngeal glands, which may compromise the survival and foraging of this pollinator.

Hypergravity Impact on Fertility of Apis mellifera carnica Queens – Case Study

The launch is considered the most stressful rocket flight stage due to the hypergravity occurrences. The possibility of using honey bees (Apis mellifera) as the extraterrestrial pollinator depends on their ability to reproduce correctly after experiencing hypergravity. The described study aims to verify the impact of a launching rocket’s acceleration on honey bee queen’s egg-laying behavior. Four artificially inseminated A. mellifera carnica queens were placed in the Human Training Centrifuge and given to the acceleration pattern of the launching Soyuz rocket. Next, the data on the number of food stores, eggs, larvae, and worker and drone pupae were collected from the test and control hives using the modified Liebefeld method. The pilot study results imply that accelerated queen’s egg-laying behavior may change twofold: limiting or maximizing the number of laid eggs, with the control queen egg-laying rate remaining stable for all samples. The number of drone pupae is greater for the test sample colonies, with its earlier appearance in the hive. No impact on overwintering success was observed. Authors indicate limitations of the results and a need to continue the study to verify the occurrence of anomalies potentially related to the examined factor.

Pesticide risk during commercial apple pollination is greater for honeybees than other managed and wild bees

Abstract Most pesticide research has focussed on risk to managed honeybees, but other managed and wild bees are also exposed to pesticides. Critically, we know little about the magnitude and sources of risk to honeybees compared with other bees during crop pollination. To compare pesticide exposure and risk across wild and managed bees, we sampled the main bee groups present during bloom in 20 apple orchards, including managed honeybees (Apis mellifera), managed bumblebee workers (Bombus impatiens), wild mining bees (Andrena spp. and Andrena [Melandrena] spp.), bumblebee foundress queens (Bombus impatiens) and eastern carpenter bees (Xylocopa virginica). We screened all bees for 92 pesticides and computed a Risk Quotient using available toxicity data (honeybee LD50s), adjusting for differences in toxicity known to scale with body mass. To gain insight into exposure origin, we compared residues in bees to those in focal orchard apple and dandelion flowers. Nearly all bee samples contained pesticides (95%), with the average contamination level ranging from 7.1 ± 2.8 parts per billion (ppb) in B. impatiens workers to 388.4 ± 146.2 ppb in Andrena. Exposure profiles were similar for all bees except A. mellifera, whose unique exposure profile included high levels of the neonicotinoid insecticide thiamethoxam. All bee groups except wild B. impatiens queens had at least one sample exceeding a US Environmental Protection Agency or European Food Safety Authority exposure level of concern. Apis mellifera experienced significantly greater risk than other bee groups, with 63% and 81% of samples exceeding an acute or chronic exposure level of concern, respectively. Risk to honeybees was driven primarily by high thiamethoxam levels not found in focal orchard flowers and likely originating outside the orchard. Synthesis and applications: We find that pesticide exposure and risk differ between honeybees and other managed and wild bees during apple pollination. Furthermore, pesticide exposure is a landscape‐scale phenomenon and therefore measures to reduce exposure must consider the surroundings beyond focal farms. Limiting orchard sprays, while reducing on‐farm exposures, will not protect far‐foraging bees from off‐farm exposures such as thiamethoxam, which we hypothesize is coming from nearby seed‐treated corn fields planted during apple bloom.

Prevalence and infection intensity of honey bee (Apis mellifera) viral diseases in six regions of the state of Jalisco, Mexico

El estado de Jalisco es un importante productor de miel de abejas en México. Sin embargo, no existe información sobre las virosis que afectan a las abejas melíferas (Apis mellifera) en las diferentes regiones apícolas de la entidad. El objetivo de este estudio fue determinar la prevalencia e intensidad de cuatro enfermedades virales de Apis mellifera durante la primavera, en seis regiones de Jalisco. Se analizaron abejas de 79 colonias, de las cuales, el 66 y 38 % fueron positivas al virus de la celda real negra (VCRN) y al virus de las alas deformes (VAD), respectivamente. Dos virosis no fueron detectadas, las causadas por el virus de la parálisis aguda Israelí (VPAI) y la causada por el virus de la parálisis crónica (VPC). Los niveles de infección fueron relativamente bajos para el VCRN, pero elevados para el VAD, cuyas infecciones fueron 8,000 veces más altas que las del VCRN. La prevalencia del VAD fue significativamente más alta en las regiones de los Altos, Centro y Sur, mientras que para el VCRN no hubo diferencias entre regiones. Para la intensidad de infecciones, no hubo diferencias entre regiones para el VAD, pero sí para el VCRN. Las regiones con niveles de infección más altos fueron la Sur y Centro. Se recomienda realizar muestreos en otras épocas del año para detectar posibles efectos estacionales de las virosis de las abejas y diseñar estrategias para su control.   

Apirhabdus apintestini gen. nov., sp. nov., a member of a novel genus of the family Enterobacteriaceae, isolated from the gut of the western honey bee Apis mellifera.

A Gram-negative, motile, rod-shaped bacterial strain, CA-0114T, was isolated from the midgut of a western honey bee, Apis mellifera. The isolate exhibited ≤96.43 % 16S rRNA gene sequence identity (1540 bp) to members of the families Enterobacteriaceae and Erwiniaceae. Phylogenetic trees based on genome blast distance phylogeny and concatenated protein sequences encoded by conserved genes atpD, fusA, gyrB, infB, leuS, pyrG and rpoB separated the isolate from other genera forming a distinct lineage in the Enterobacteriaceae. In both trees, the closest relatives were Tenebrionicola larvae YMB-R21T and Tenebrionibacter intestinalis BIT-L3T, which were isolated previously from Tenebrio molitor L., a plastic-eating mealworm. Digital DNA-DNA hybridization, orthologous average nucleotide identity and average amino acid identity values between strain CA-0114T and the closest related members within the Enterobacteriaceae were ≤23.1, 75.45 and 76.04 %, respectively. The complete genome of strain CA-0114T was 4 451669 bp with a G+C content of 52.12 mol%. Notably, the apparent inability of strain CA-0114T to ferment d-glucose, inositol and l-rhamnose in the API 20E system is unique among closely related members of the Enterobacteriaceae. Based on the results obtained through genotypic and phenotypic analysis, we propose that strain CA-0114T represents a novel species and genus within the family Enterobacteriaceae, for which we propose the name Apirhabdus apintestini gen. nov., sp. nov. (type strain CA-0114T=ATCC TSD-396T=DSM 116385T).

Interaction of acetamiprid, Varroa destructor, and Nosema ceranae in honey bees

Health of honey bees is threatened by a variety of stressors, including pesticides and parasites. Here, we investigated effects of acetamiprid, Varroa destructor, and Nosema ceranae, which act either alone or in combination. Our results suggested that interaction between the three factors was additive, with survival risk increasing as the number of stressors increased. Although exposure to 150 μg/L acetamiprid alone did not negatively impact honey bee survival, it caused severe damage to midgut tissue. Among the three stressors, V. destructor posed the greatest threat to honey bee survival, and N. ceranae exacerbated intestinal damage and increased thickness of the midgut wall. Transcriptomic analysis indicated that different combinations of stressors elicited specific gene expression responses in honey bees, and genes involved in energy metabolism, immunity, and detoxification were altered in response to multiple stressor combinations. Additionally, genes associated with Toll and Imd signalling, tyrosine metabolism, and phototransduction pathway were significantly suppressed in response to different combinations of multiple stressors. This study enhances our understanding of the adaptation mechanisms to multiple stressors and aids in development of suitable protective measures for honey bees. Environmental ImplicationWe believe our study is environmentally relevant for the following reasons: This study investigates combined effects of pesticide, Varroa destructor, and Nosema ceranae. These stressors are known to pose a threat to long-term survival of honey bees (Apis mellifera) and stability of the ecosystems. The research provides valuable insights into the adaptive mechanisms of honey bees in response to multiple stressors and developing effective conservation strategies. Further research can identify traits that promote honey bee survival in the face of future challenges from multiple stressors to maintain the overall stability of environment.

Melittin—The principal toxin of honeybee venom—Is also produced in the honeybee fat body

Melittin is a powerful toxin present in honeybee venom that is active in a wide range of animals, from insects to humans. Melittin exerts numerous biological, toxicological, and pharmacological effects, the most important of which is destruction of the cell membrane. The phospholipase activity of melittin and its ability to activate phospholipases in the venom contribute to these actions. Using analytical methods, we discovered that the honeybee Apis mellifera produces melittin not only in the venom gland but also in its fat body cells, which remain resistant to this toxin's effects. We suggest that melittin acts as an anti-bacterial agent, since its gene expression is significantly upregulated when honeybees are infected with Escherichia coli and Listeria monocytogenes bacteria; additionally, melittin effectively kills these bacteria in the disc diffusion test. We hypothesize that the chemical and physicochemical properties of the melittin molecule (hydrophilicity, lipophilicity, and capacity to form tetramers) in combination with reactive conditions (melittin concentration, salt concentration, pH, and temperature) are responsible for the targeted destruction of bacterial cells and apparent tolerance towards own tissue cells. Considering that melittin is an important current and, importantly, potential broad-spectrum medication, a thorough understanding of the observed phenomena may significantly increase its use in clinical practice.

Host Barriers Limit Viral Spread in a Spillover Host: A Study of Deformed Wing Virus in the Bumblebee Bombus terrestris.

The transmission of pathogens from reservoir to recipient host species, termed pathogen spillover, can profoundly impact plant, animal, and public health. However, why some pathogens lead to disease emergence in a novel species while others fail to establish or do not elicit disease is often poorly understood. There is strong evidence that deformed wing virus (DWV), an (+)ssRNA virus, spills over from its reservoir host, the honeybee Apis mellifera, into the bumblebee Bombus terrestris. However, the low impact of DWV on B. terrestris in laboratory experiments suggests host barriers to virus spread in this recipient host. To investigate potential host barriers, we followed the spread of DWV genotype B (DWV-B) through a host's body using RT-PCR after experimental transmission to bumblebees in comparison to honeybees. Inoculation was per os, mimicking food-borne transmission, or by injection into the bee's haemocoel, mimicking vector-based transmission. In honeybees, DWV-B was present in both honeybee faeces and haemolymph within 3 days of inoculation per os or by injection. In contrast, DWV-B was not detected in B. terrestris haemolymph after inoculation per os, suggesting a gut barrier that hinders DWV-B's spread through the body of a B. terrestris. DWV-B was, however, detected in B. terrestris faeces after injection and feeding, albeit at a lower abundance than that observed for A. mellifera, suggesting that B. terrestris sheds less DWV-B than A. mellifera in faeces when infected. Barriers to viral spread in B. terrestris following oral infection may limit DWV's impact on this spillover host and reduce its contribution to the community epidemiology of DWV.

The use of honey bees (Apis mellifera L.) to monitor airborne particulate matter and assess health effects on pollinators.

The honey bee Apis mellifera has long been recognized as an ideal bioindicator for environmental pollution. These insects are exposed to pollutants during their foraging activities, making them effective samplers of environmental contaminants, including heavy metals, pesticides, radionuclides, and volatile organic compounds. Recently, it has been demonstrated that honey bees can be a valuable tool for monitoring and studying airborne PM pollution, a complex mixture of particles suspended in the air, known to have detrimental effects on human health. Airborne particles attached to the bees can be characterised for their morphology, size, and chemical composition using a scanning electron microscopy coupled with X-ray spectroscopy, thus providing key information on the emission sources of the particles, their environmental fate, and the potential to elicit inflammatory injury, oxidative damage, and other health effects in living organisms. Here, we present a comprehensive summary of the studies involving the use of honey bees to monitor airborne PM, including the limits of this approach and possible perspectives. The use of honey bees as a model organism for ecotoxicological studies involving pollutant PM is also presented and discussed, further highlighting the role of the bees as a cornerstone of human, animal, and environmental health, according to the principles of the "One Health" approach.

Nurse bees regulate the larval nutrition of developing workers (Apis mellifera) whenfeeding on various pollen types.

Nutrition has been identified as a key driver of colony health and productivity. Yet, in honey bees, relatively little is known about how the vast variety of natural pollen sources impact larval development. The impact of the nutritional quality of 4 naturally occurring pollen sources, of importance to the Western Australian beekeeping industry, was tested on honey bee (Apis mellifera L.) development. Bee packages consisting of 800g of bees and a mated sister queen were assigned to 40 nucleus hives and randomly allocated to one of the 4 feed treatments (10 colonies each) of marri (Corymbia calophylla Lindl.), jarrah (Eucalyptus marginata Sm.), clover (Trifolium repens L.), and canola (Brassica napus L.) pollen. Emerging bees were collected once the first bees started hatching on the assigned feed sources. Newly emerging bees were weighed individually, and body composition was measured in batches according to the feed treatment groups. Food consumption was recorded for the duration of the experiment. Nurse bees successfully raised young adult workers from the larval stage until emergence when fed with one of 4 pollen patties with different nutritional qualities. There was no difference in the body composition or weight of emerging bees fed on the different pollen types. However, the body weight of bees increased over time, most likely related to colony size and structure. With the type of pollen patties having little impact on larval development, the availability of pollen may be more important than its composition, providing bees have access to all essential nutrients.

Revealing Changes in Ovarian and Hemolymphatic Metabolites Using Widely Targeted Metabolomics between Newly Emerged and Laying Queens of Honeybee (Apis mellifera).

The queen bee is a central and pivotal figure within the colony, serving as the sole fertile female responsible for its reproduction. The queen possesses an open circulatory system, with her ovaries immersed in hemolymph. A continuous and intricate transportation and interchange of substances exist between the ovaries and hemolymph of queen bees. To determine the characteristic metabolites in the hemolymph and ovary, as well as understand how their rapid metabolism contributes to the process of egg-laying by queens, we reared Apis mellifera queens from three different age groups: newly emerged queen (NEQ), newly laying queen (NLQ), and old laying queen (OLQ). Using widely targeted metabolomics, our study revealed that the laying queen (NLQ and OLQ) exhibited faster fatty acid metabolism, up-regulated expression of antioxidants, and significant depletion of amino acids compared to the NEQ. This study revealed that the levels of carnitine and antioxidants (GSH, 2-O-α-D-glucopyranosyl-L-ascorbic acid, L-ascorbic acid 2-phosphate, etc.) in the NLQ and OLQ were significantly higher compared to NEQ. However, most of the differentially expressed amino acids, such as L-tryptophan, L-tyrosine, L-aspartic acid, etc., detected in NLQ and OLQ were down-regulated compared to the NEQ. Following egg-laying, pathways in the queens change significantly, e.g., Tryptophan metabolism, Tyrosine metabolism, cAMP signaling pathway, etc. Our results suggest that carnitine and antioxidants work together to maintain the redox balance of the queen. Additionally, various amino acids are responsible for maintaining the queen's egg production.

The Influence of Brood Survival Rates on the Behavioral Patterns, Economic Aspects and Resistant Traits of Italian Honeybee Apis mellifera L. Reared on Indian Mustard in Bihar

To better understand Apis mellifera foraging behaviour for mustard, this study examined how brood survival rates affected the species behavioural, economic and resistant traits. Out of 10 colonies used for the study, the results showed that 1 colony had a low rate of brood survival, 3 colonies had a medium rate, and 6 colonies had a high rate. In the experiment on scattered brood patterns, poor, medium and excellent brood patterns were found in 1, 2, and 7 colonies, respectively. According to the data on bee activity collected, the majority of bees were observed leaving between the hours of 9:00 and 10:00 and arriving between the hours of 16:00 and 17:00. According to the economic attributes data, colonies with high and moderate brood survival rates had more honey and pollen reserves. According to the hygienic behaviour results, colonies with high and moderate brood survival rates had 100 per cent efficiency while colonies with low brood survival rates had the lowest efficiency at 76 per cent.

Factors Affecting Honey Bee (Apis mellifera L.) Health as Perceived by South Georgia Commercial Beekeepers

Honey bees (Apis mellifera L.) are critical to the pollination of many crops. Bee-oriented organizations survey beekeepers annually to gather information about colony loss and contributing factors. Unfortunately, these surveys provide insufficient data from sideliner and commercial beekeepers in Georgia. Through a survey to better understand South Georgia beekeepers’ perceptions of honey bee health, this project engaged a previously underrepresented group of sideliner and commercial beekeepers.

Molecular detection of Lotmaria passim in intestine of Japanese honeybees (Apis cerana japonica)

ABSTRACTCrithidia mellificae and Lotmaria passim are trypanosomatids that infect honeybees, and many studies suggest that both parasites contribute to the decline in the honeybee population. The Japanese honeybee (Apis cerana japonica) is a native honeybee that inhabits various areas of Japan and is one of the most important pollinators. Both parasites have been well studied in the western honeybee (Apis mellifera) but have been poorly investigated in A. c. japonica. The present study investigated the presence of C. mellificae and L. passim in one feral and six managed A. c. japonica colonies at three different institutes. Five out of seven colonies were polymerase chain reaction‐positive for L. passim (71.4%); however, C. mellificae was not detected. Four of the five colonies were positive in both the midgut and hindgut, but one was positive only in the hindgut. A homology search and phylogenetic tree of the glycosomal glyceraldehyde‐3‐phosphate dehydrogenase (gGAPDH) and cytochrome b (Cytb) genes obtained in this study indicated that the sequences belonged to L. passim. The results of this study emphasize concerns regarding the health of this ecologically important pollinator and the importance of further investigation of the prevalence of L. passim in A. c. japonica in a wider area of Japan.