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On-site DNA Detection of Trypanosomatid Parasites and Nosema ceranae Through Alkaline Lysis Coupled to RPA/CRISPR/Cas12a System
Published on: July 18, 2025
Microsporidia Nosema spp. - obligate bee parasites are transmitted by air
Aneta Sulborska1, Beata Horecka2, Malgorzata Cebrat3
1Department of Botany, University of Life Sciences, Akademicka 15, 20-950, Lublin, Poland.
Abstract:
Microsporidia Nosema are transferred among bees via the faecal-oral route. Nosema spp. spores have been detected on flowers and transferred to hives along with the bee pollen. The aim of the present study was to determine whether Nosema microsporidia are transferred by air in an apiary, in a control area (without the presence of bee colonies), and/or in a laboratory during cage experiments with artificially infected bees. The novel way of transmission by air was investigated by the volumetric method using a Hirst-type aerobiological sampler located on the ground in the apiary, in the Botanical Garden and on the laboratory floor. Concurrently, the mean rate of Nosema infections in the foragers in the apiary was estimated with the Bürker haemocytometer method. Spore-trapping tapes were imaged by means of light microscopy, Nomarski interference contrast microscopy and scanning electron microscopy. The highest concentration of Nosema spores per 1m3 of air (4.65) was recorded in August, while the lowest concentration (2.89) was noted in July. This was confirmed by a Real-Time PCR analysis. The presence of N. apis as well as N. ceranae was detected in each of the tested tapes from the apiary. The average copy number of N. apis was estimated at 14.4 × 104 copies per 1 cm2 of the tape; whereas the number of N. ceranae was 2.24 × 104 copies per tape per 1 cm2. The results indicate that Nosema microsporidia were transferred by the wind in the apiary, but not in the Botanical Garden and laboratory by air. This was confirmed by genetic analyses. DNA from immobilised biological material was isolated and subjected to a PCR to detect the Nosema species. A fragment of the 16S rRNA gene, characteristic of Nosema apis and N. ceranae, was detected. Our research adds knowledge about the transfer of Nosema spp. microsporidia in the natural environment and indicates the season associated with the greatest risk of a bee colony infection with Nosema spp.
Insights
Airborne Nosema microsporidia transmission was confirmed in apiaries, posing a risk to bee colonies. This study highlights wind as a vector for Nosema spp. spores, particularly in August, impacting bee health.
Area of Science:
- Environmental microbiology
- Apiculture science
- Insect pathology
Background:
- Microsporidia Nosema are known bee pathogens transmitted via faecal-oral routes.
- Nosema spores can contaminate flowers and pollen, leading to hive infections.
- Understanding transmission routes is crucial for managing bee diseases.
Purpose of the Study:
- To investigate the potential for airborne transmission of Nosema microsporidia in apiaries.
- To compare airborne Nosema presence in apiaries versus control areas (Botanical Garden, laboratory).
- To identify the primary Nosema species involved and seasonal infection risks.
Main Methods:
- Utilized a Hirst-type aerobiological sampler for volumetric spore collection in apiaries and control sites.
- Employed light, Nomarski interference contrast, and scanning electron microscopy for spore imaging.
- Conducted Real-Time PCR and genetic analysis (16S rRNA gene) to detect and quantify Nosema species (N. apis, N. ceranae).
- Estimated Nosema infection rates in foragers using the Bürker haemocytometer method.
Main Results:
- Nosema spores were detected in the air of the apiary, with the highest concentration (4.65 spores/m³) in August.
- Airborne Nosema transmission was not observed in the Botanical Garden or laboratory settings.
- Both Nosema apis and Nosema ceranae were identified in apiary air samples via PCR.
- Significant spore concentrations (N. apis: 14.4 × 10⁴ copies/cm², N. ceranae: 2.24 × 10⁴ copies/cm²) were quantified.
Conclusions:
- Wind facilitates the airborne transmission of Nosema microsporidia within apiaries.
- August presents the highest risk period for Nosema spp. infection spread via air.
- This finding adds crucial knowledge to Nosema spp. transmission dynamics in natural environments.
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